The epidemiology of hospitalised acute kidney injury not requiring dialysis in England from 1998 to 2013: retrospective analysis of hospital episode statistics.

AIMS: Epidemiology studies of acute kidney injury (AKI) have focused on cases requiring dialysis but those not requiring dialysis represent the majority. To address this gap, we interrogated hospital episode statistics (HES) to investigate population trends in temporal epidemiology of AKI not requiring dialysis between 1998 and 2013. METHODOLOGY: In this retrospective observational study of HES data covering the entire English National Health Service, we identified 1,136,167 AKI events, not requiring dialysis, diagnosed between 1998 and 2013. We explored the effect of age, gender, ethnicity, Charlson's comorbidity score (CCS), method of admission, diagnosis period and AKI in diagnosis codes on temporal changes in the incidence and case-fatality of AKI with specific examination of its predictors. RESULT: The incidence of AKI increased from 15,463 cases (317 pmp) in 1998-1999 to 213,700 cases (3995 pmp) in 2012-2013. There was increase in proportion of people over 75 years from 51.1% in 1998-1999 to 63.4% in 2012-2013. Overall unadjusted case-fatality decreased from 42.3% in 1998-2003 to 27.1% in 2008-2013, p < 0.001. Compared with 1998-2003, the multivariable adjusted odds ratio for death was 0.64 in 2003-2008 (95% CI 0.63-0.65) and 0.35 in 2008-2013 (95% CI 0.34-0.35). Odds for death were higher for patients over 85 years (2.93; 95% CI 2.89-2.97), CCS of more than five (2.75; 95% CI 2.71-2.79), emergency admissions (2.14; 95% CI 2.09-2.18) and AKI in the secondary diagnosis code (1.35; 95% CI 1.33-1.36) and AKI in other diagnoses codes (2.17; 95% CI 2.15-2.20). CONCLUSIONS: In England, the incidence of AKI not requiring dialysis has increased and case-fatality has decreased over last 15 years. Efforts to reduce the incidence of AKI and improve survival should focus on elderly people, emergency admissions and those with multi-morbidity.

Creation of an arteriovenous fistula is associated with significant acute local and systemic changes in microvascular function.

BACKGROUND: Native arteriovenous fistulae (AVF) are the vascular access of choice for haemodialysis. The consequences of AVF formation on microvascular function, locally or systemically, are unknown. METHODS: We recruited 43 predialysis patients undergoing AVF formation. Patients were studied 2 weeks prior to the planned AVF operation and 2 weeks postoperatively. Thirteen patients with failed AVF were subsequently utilised as sham controls. Laser Doppler perfusion imaging was used to measure subcutaneous microvascular blood flow. Microvascular function was assessed as an increase in perfusion in response to iontophoretic administration of vasodilatory stimuli assessing endothelial-dependent (ED) and non-endothelial-dependent (NED) vasodilatation. RESULTS: Patients with successful AVF formation had a significantly reduced ED vasodilatation in the fistula arm (-36 ± 46%, p < 0.001). Only NED vasodilatation was significantly reduced in the non-fistula arm (23 ± 40%, p = 0.01). Patients who had an unsuccessful AVF operation exhibited no recordable changes. CONCLUSIONS: Formation of an AVF was associated with local and remote changes in microcirculation. Further assessments are underway to examine the contributions of local shear stress, vasoreactive substances and the autonomic responses. Although the clinical significance of these findings is not yet clear, it is intriguing that AVF formation is associated with such widespread and profound changes in microperfusion.

Biofeedback controlled hemodialysis (BF-HD) reduces symptoms and increases both hemodynamic tolerability and dialysis adequacy in non-hypotension prone stable patients.

BACKGROUND: It is now possible to link relative blood volume (RBV) measurements to a software loop designed to actuate a biofeedback response. This allows changes in RBV to determine constant alterations in both ultrafiltration rate and dialysate conductivity. RBV, plasma sodium and weight loss are driven throughout the treatment to achieve the best compromise. This system has been demonstrated to markedly reduce intradialytic hypotension in unstable patients. We have applied this treatment to stable, non-hypotension prone HD patients and report on the short-term outcomes. METHODS: We prospectively studied all 15 patients in a dedicated 4-station minimal care treatment area. Patients were studied for 3 weeks of standard HD, to understand the morphology and response to RBV in that individual. BF-HD was then instituted for a similar period (after a 2-week optimization period). Dialysis adequacy was assessed with equilibrated Kt/V measurements and urea mass removed in spent dialysate. RESULTS: We studied 263 treatment sessions. There was a reduction in symptomatic episodes (per patient over 3 weeks) from 3 +/- 0.5 (0-9) to 0.13 +/- 0.13 (0-2) with BF-HD, p < 0.001. Reductions in systolic BP > 40% fell from 1.4 +/- 0.4 (0-4) to 0.46 +/- 0.16 (0-2). Episodes of RBV falling > 10% fell from 6.3 +/- 0.85 (1-13) to 1.13 +/- 0.27 (0-4) with BF-HD, p < 0.001. Interdialytic weight gains fell from 2.08 +/- 0.05 (0.35-3.8) kg to 1.82 +/- 0.06 (0-3.7) kg, p = 0.009. Equilibrated Kt/V increased from 1.01 +/- 0.03 (0.61-1.35) to 1.13 +/- 0.03 (0.7-1.5), p = 0.01, and mass removed of urea increased from 24.9 +/- 3 (12.8-45) g to 32.7 +/- 1.9 (17.3-48.5) g. CONCLUSIONS: This is the first report of BF-HD increasing tolerability, reducing interdialytic fluid gains and enhancing urea clearance in non-hypotension prone chronic HD patients. These data suggest that the previously reported associated benefits of BF-HD may be applicable to the majority of HD patients.

Characterization of treatment dose delivered by albumin dialysis in the treatment of acute renal failure associated with severe hepatic dysfunction.

BACKGROUND: Acute liver cell failure (ALCF) commonly results in death and when complicated by acute renal failure (ARF), the mortality approaches 90%. Albumin dialysis allows partial replacement of some of the liver's excretory functions. The molecular absorbents recirculating system (MARS) has been recently introduced to provide this therapy. Thus allowing bridging to transplantation or hepatic regeneration. We have attempted to define the degree of "uremic" dialysis that this system can deliver as well as characterizing the dose of "hepatic" treatment, using a similar approach to solute remove as applied to assessing hemodialysis adequacy. As a secondary issue we also report on the clinical outcomes of this group of patients. METHOD: We treated 7 patients with ALCF and acute renal failure (6 of the patients having a formal diagnosis of hepatorenal syndrome), aiming to deliver a 5 treatment consecutive course consisting of 8 hours of albumin dialysis using the MARS monitor, combined with hemodialysis. Clinical and biochemical outcomes were assessed, and dialysis adequacy measured using urea reduction ratios, calculated Kt/V and measured Kt/V (using ionic dialysance). Treatment dose, with respect to the highly protein bound and lipophilic toxins that accumulate in hepatic failure, was assessed by calculating the bilirubin reduction ratio and percentage reduction in plasma ammonia and total bile acids. RESULTS: All of the patients had a degree of biochemical improvement with albumin dialysis. Urine output increased and the degree of encephalopathy improved. Mean bilirubin fell from 612 +/- 105.5 micromol/l (range 165.6 - 1,024 micromol/l) to 370.4 +/- 49.7 micromol/l (range 190.4 - 569.2 micromol/l), ALT reduced from 3,280 +/- 2,266 IU/l (range 40 - 18,876) to 639 +/- 230 IU/l (range 33 - 1677). Hepatic synthetic function improved with INR falling from 4.1 +/- 0.5 (range 2.1 - 6.4) to 2.8 +/- 0.6 (range 1.4 - 5.5). Plasma ammonia was reduced, falling from 162.4 +/- 15.4 (range 131.1 - 191.9 micromol/l) to 73.1 +/- 15 micromol/l (range 45.6 - 106.4 micromol/l). Bile acid levels fell from 132 +/- 10.2 micromol/l (range 110.7 - 155.8 micromol/l) to 36.9 +/- 6.1 micromol/l (range 24.6 49.6 micromol/l). The mean urea reduction ratio (URR) was 58.4 +/- 3.2% (range 39 - 76%). Mean Kt/V as assessed by ionic dialysance was 1.7 +/- 0.01 (range 0.8-2.4). Mean bilirubin reduction ratio (BRR) was 28.6 +/- 1.4% (range 12.5 - 39%). BRR was proportional to both URR and Kt/V. BRR was also proportional to the percentage reduction of ammonia and bile acid levels. Three of the 7 patients survived to be discharged from hospital and 4 died. CONCLUSION: Albumin dialysis appears capable of improving the outcome in patients with ALCF and hepatorenal syndrome. Eight-hour intermittent treatments with the MARS system in combination with hemodialysis deliver an adequate dose of dialysis with respect to urea. BRR may be an appropriate tool to allow further quantitative and comparative study of this technique.

Simultaneous non-surgical removal and insertion of peritoneal dialysis catheters for defective drainage: a bedside, day case technique.

BACKGROUND: Outflow failure of peritoneal dialysis catheters is a commonly encountered problem. It may be possible to reposition the catheter by a variety of means, but this can be problematical and has variable long-term success. Commonly surgical catheter exchange is utilized, entailing inconvenience, expense and often, a reliance on temporary hemodialysis. We describe a technique allowing exchange of poorly functioning catheters with a relatively simple outpatient/day case percutaneous technique, allowing the continuation of peritoneal dialysis. METHODS: We report percutaneous exchange of 25 peritoneal dialysis catheters in 21 patients. The exchanges were performed under local anesthesia with a degree of sedation (if required). It involved the dissection down the distal cuff of the catheter and mobilization of the catheter below it. This was followed by division of the catheter, allowing passage of a guide wire into the peritoneal cavity and insertion of a further peel away sheath and insertion of a new catheter. The new catheter was tunneled out of the existing exit site after removal of the extraperitoneal portion of the old catheter by traction. RESULTS: Outflow failure was associated with fecal loading and malposition of the catheter in 14 out of the 21 patients. Exchange of catheter was successful in all the patients with good pelvic positioning of the replacement catheter in all but 1 of the cases. The mean period until the reinstitution of peritoneal dialysis was 5.1 days (range 0-14 days). Temporary hemodialysis was not required for any of the patients. One patient exhibited a small leak of peritoneal dialysis fluid after insertion, but this had spontaneously resolved within 6 days. Protracted satisfactory function of the peritoneal dialysis catheters was obtained in all but 1 of the patients (mean follow-up 51 weeks, range 11-73 weeks). CONCLUSIONS: We conclude that exchange of peritoneal dialysis catheters for problems with dialysate drainage, utilizing a non-invasive percutaneous technique is both effective and safe.

Long-term use of 1.1% amino acid dialysis solution in hypoalbuminemic continuous ambulatory peritoneal dialysis patients.

INTRODUCTION: Malnutrition is a common problem in patients treated with continuous ambulatory peritoneal dialysis (CAPD). Hypoalbuminemia in CAPD patients is an independent risk factor for death and is associated with malnutrition. Previous short-term studies have examined the use of amino acid based PD solutions in terms of albumin levels and anthropometric changes, but not clinical outcome. We report on the extended use of 1.1% amino acid based peritoneal dialysis solution (Nutrineal) and have assessed clinical utility in terms of nutrition, biochemical indices, dialysis adequacy and clinical outcomes. METHODS: The effect of Nutrineal was studied retrospectively in 22 patients during the past 30 months. All patients had an albumin level of < 35 g/l prior to commencing Nutrineal, and had either a protein intake < 1.2 g/kg or weight loss of > 5% in the previous 3 months. 19 of the 22 patients underwent an 8-week trial of oral nutritional supplements with no improvement in serum albumin level. Albumin level, normalized protein catabolic rate, weight, Kt/V and creatinine clearance were assessed for all patients prior to Nutrineal and at the end of the study period. RESULTS: The mean time on Nutrineal therapy was 13.6 months (range 6-26 months). There were no reported side effects of the treatment. There was an average of 1 episode of peritonitis per 23 treatment months, and only 1 patient died (4% annually adjusted mortality cf 8.9% on the peritoneal dialysis program as a whole). There was a significant increase in albumin level from 22.45 +/- 0.97 range 14-33 g/l to 25.68 +/- 1.159 range 16-35 g/l (p = 0.0036). Normalized protein catabolic rate increased significantly, from 0.898 +/- 0.053 to 1.085 +/- 0.056 g/kg/day (p = 0.0057). Weight decreased slightly although this did not reach statistical significance. Kt/V and creatinine clearance both decreased significantly, but remained within the adequate range in > 80% of the patients. There was no significant change in residual renal function (mean residual creatinine clearance 3.8 +/- 0.59 ml/min at the start of the study period, cf 3.4 +/- 0.61 ml/min at the end). CONCLUSION: These data suggest that Nutrineal can be used safely and effectively for an extended period of time. Such use is associated with a low mortality rate and a low peritonitis rate, although dialysis adequacy is compromised to a degree.

Steroid and cyclophosphamide therapy for IgA nephropathy associated with crescenteric change: an effective treatment.

BACKGROUND: IgA nephropathy is the most common form of idiopathic glomerulonephritis. There is no current consensus on treatment for this condition. We report on the effect of immunosuppression with corticosteroids and cyclophosphamide for the treatment of IgA nephropathy associated with crescenteric change. METHODS: The effect of oral prednisolone (0.8 mg/kg initially, reducing to 0.4 mg/kg after 4 weeks) and cyclophosphamide (1.5 mg/kg) given until a plateau of response was obtained was studied in 9 patients with IgA nephropathy associated with severe inflammatory change and crescents. The initial diagnostic renal biopsies of these patients revealed 25-70% of the glomeruli effected with active cellular crescents. When response to therapy, plateaued cyclophosphamide was discontinued and prednisolone reduced from 0.4 mg/kg. Follow-up renal biopsy was performed in 8 of the 9 patients. Patients were maintained on prednisolone (5- 7.5 mg) and azathioprine (1 mg/kg) for further 2 years. RESULTS: The mean time until discontinuation of cyclophosphamide was 17.8 weeks (+/-1.23, range 12-25 weeks). There were no serious complications of therapy. There was an improvement in renal function in all patients with serum creatinine falling from a mean of 149.6+/-16.5, range 81-227 micromol/l to 116.4+/-8.6, range 80-158 micromol/l, p=0.01. Creatinine clearance improved from a mean of 57.1+/-9.9, range 21-104 ml/min to 87.2+/-10.1, range 39-125 ml/min, p=0.004. 24-hour urinary total protein fell over the same time m period from a mean of 4.54+/-1.1, range 1.0-11.27 g to 1.2+/-0.27, range 0.01-2.65 g, p=0.004. There were no significant differences in blood pressure during this time. Repeat renal biopsies showed significant degrees of histological improvement with healing of crescents and a reduction in acute inflammatory change in all except one patient. The mean period of follow-up after cessation of cyclophosphamide was 17.4+/-2.8 months, range 10-36 months. There was no significant change over this period in serum creatinine, creatinine clearance or urinary protein losses. CONCLUSION: These data suggest that IgA nephropathy associated with severe inflammatory and crescenteric change can be effectively and safely treated with a low-cost regime based on oral corticosteroids and cyclophosphamide tailored to a plateau of treatment response in individual patients.

Contractile dysfunction of isolated ventricular myocytes in experimental uraemia.

In order to clarify the mechanism underlying impaired cardiac performance in uraemia, the contractile function of isolated cardiac myocytes from chronically uraemic and control rats has been compared. Rats were made uraemic by sub-total nephrectomy in a two-stage surgical procedure, and left for 4 weeks. Sham-operated controls were prepared at the same time. Animals were pairfed, and final body weights were not significantly different between the groups. Ventricular myocytes were isolated and their contraction amplitude and velocity were measured using a video-based edge-detection system. Contraction was depressed in myocytes from uraemic animals, with contraction amplitude in maximum Ca2+ reduced from 16.3 +/- 0.6% shortening, to 13.0 +/- 0.8% (p < 0.01, n = 10 animals for each group). There was a concomitant decrease in the velocity of shortening (5.6 +/- 0.4 vs. 3.9 +/- 0.5 micron s-1 change in sarcomere length, p < 0.02) and of relaxation (4.6 +/- 0.4 vs. 3.2 +/- 0.4 micron s-1 p < 0.02). Similar depression was seen at lower perfusate Ca2+ concentrations (1-2 mM) and the EC50 for Ca2+ was unchanged. The response to beta-adrenoceptor stimulation was decreased by the same magnitude as that to Ca2+, with no change in the EC50 for isoproterenol or the ratio of maximum response to isoproterenol or to Ca2+ in the same cell (isoproterenol/Ca2+ ratio). There was no shift in the myosin isozyme composition in uraemic cells, with both groups showing a heterogeneous V1/V2/V3 pattern. We conclude that chronic uraemia is associated with a depression of contractile function in the isolated myocyte but no shift in myosin isoforms or specific beta-adrenoceptor desensitisation.

Recombinant erythropoietin increases blood pressure in experimental hypertension and uraemia without change in vascular cytosolic calcium.

The mechanism of erythropoietin-induced hypertension in dialysis patients is unclear. Intracellular calcium ([Ca2+]i) may be altered in both hypertension and uraemia, and the effects of both uraemia and r-HuEPO on vascular smooth muscle [Ca2+]i and blood pressure (BP) in Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR) were therefore studied. Male WKY and SHR underwent partial nephrectomy or sham operation. Three weeks later a 28-day period of treatment with either r-HuEPO 100 U/kg, s.c., 3 times/week or buffer was commenced (n = 10-12 for each subgroup). BP was measured weekly, by noninvasive Doppler tail-cuff assessment. [Ca2+]i was measured following loading with fura-2 in pooled, primary aortic vascular smooth muscle cells (VSMC). Serum urea and creatinine rose 3- to 4-fold after partial nephrectomy. Treatment with r-HuEPO did not change renal function further in either uraemic or control WKY or SHR. Haemoglobin increased in both non-uraemic WKY (16.2-20.3 g/dl) and SHR (16.4-20.5 g/dl) and uraemic animals (WKY 13.9-20.9; SHR 13.8-18.8 g/dl; p < 0.01 for all changes) following 4 weeks of r-HuEPO treatment. BP was unaffected by r-HuEPO in WKY but increased in nonuraemic SHR (210-250; p < 0.01) and in uraemic SHR (224-251 mm Hg; p < 0.001) at 4 weeks. VSMC [Ca2+]i was higher in SHR than WKY (121 vs. 83 nmol/l; MANOVA p < 0.05) but no effect of uraemia or r-HuEPO on [Ca2+]i was detected. In conclusion, the hypertensive effects of r-HuEPO are augmented both in a genetic model of hypertension and in uraemia. Although VSMC [Ca2+]i was elevated in SHR, the further increase in BP induced by r-HuEPO was not associated with alterations in VSMC cytosolic calcium.

Effects of parathyroid hormone and 1,25(OH2)D3 on protein glycation in moderate uraemia.

Glucose intolerance in uraemia may be a consequence of secondary hyperparathyroidism. In this study fructosamine and glycated albumin have been used as markers of long-term glycaemic control in a group of pre-end-stage, non-diabetic uraemic patients with secondary hyperparathyroidism. The serum fructosamine level (mumol/100 g total protein) was significantly higher (p = 0.005) in uraemic patients (364 +/- 42) than in a group of 25 non-uraemic controls (332 +/- 27), but the content of glycated albumin did not differ (p > 0.05; 1.6 +/- 0.5 vs. 1.5 +/- 0.3%). In the uraemic patients, there was a significant relationship between serum 1,25-dihydroxycholecalciferol [1,25(OH2)D] (median 4.2, range 1.0-38 ng/l) and fructosamine (r = -0.66, p < 0.01; fructosamine = -2.76 1,25(OH2)D + 389), but not glycated albumin (r = -0.22, p > 0.1). No relationship existed between serum parathyroid hormone (median 15.4, range 7.0-55 pmol/l) and either glycated albumin or fructosamine (p > 0.1). In patients treated with oral calcitriol (0.25 microgram/day), significant reductions in serum parathyroid hormone after both 4 (p = 0.03) and 8 weeks (p = 0.02) and concomitant increases in serum 1,25(OH2)D (p < 0.02) after 8 weeks of treatment were not accompanied by any change in fructosamine, glycated albumin, total calcium, or ionized calcium (p > 0.05). Elevation of serum fructosamine in these patients is consistent with the impaired glucose tolerance of uraemia. The evidence presented supports a relationship between long-term glycaemic control and 1,25(OH2)D3, but not parathyroid hormone, in moderately uraemic patients with secondary hyperparathyroidism; however, serum fructosamine was not altered by treatment with calcitriol over an 8-week period.

Effects of the selective protein kinase C inhibitor Ro 31-7549 on human angiotensin II receptor desensitisation and intracellular calcium release.

The mechanism underlying type I angiotensin II (Ang II) receptor (AT1 receptor) desensitisation is unknown. Structural features suggest it may be a substrate for protein kinase C (PKC). The effects of a selective PKC inhibitor, Ro 31-7549, on receptor desensitisation were investigated in CHO cells expressing the human AT1 receptor. Desensitisation was demonstrated with respect to the calcium response to Ang II in Fura-2-loaded cells. Ro 31-7549 had no effect on desensitisation. However, pretreatment with Ro 31-7549 caused a dose-dependent reduction in calcium release from intracellular stores. PKC may therefore act at a locus distal from the receptor itself.

ACE inhibitors in non-diabetic renal disease.

In non-diabetic renal disease ACE inhibitors have brought both benefit and problems. On the one hand, they are undoubtedly effective antihypertensive agents and data suggest that they may have a beneficial role in the prevention of progression of renal disease, although further placebo controlled double blind trials of adequate duration are required. On the other hand, their widespread use in elderly patients and those with generalised atherosclerosis has increased the risk of acute renal failure when occult renovascular disease is present. Awareness among physicians of the high rate of renovascular disease in populations at risk is to be encouraged.

Modulation of platelet cytosolic calcium during erythropoietin therapy in uraemia.

Erythropoietin therapy for uraemic anaemia is associated with a high rate of hypertensive and thrombotic complications. The mechanism is unknown, but a change in cellular calcium control may be relevant to changes in blood pressure and thrombosis. Platelets were utilized as a model of vascular smooth muscle cells. The effects of erythropoietin therapy on platelet cellular calcium, assessed by fura-2, were measured in 25 patients receiving renal replacement therapy during a 6-month treatment period. Three patients failed to reach a target haemoglobin and were excluded from the analysis. Blood pressure increased in 11 of the remaining 22 subjects, eight requiring an increase in antihypertensive medication. There were no differences in cellular calcium control between the group in whom blood pressure rose and patients with stable blood pressure. Overall there was a fall of 24% in resting cytosolic calcium (baseline 69.2 +/- 5.1 to 52.5 +/- 3.0 nmol/l, P < 0.05) after 3 months of erythropoietin therapy. There was no change in the thrombin-stimulated peak response in the presence of extracellular calcium during therapy, although thrombin-stimulated intracellular release also fell at 3 months (baseline 769 +/- 61 versus 3 months 559 +/- 49 nmol/l, P < 0.01). This study suggests that intracellular free calcium control within platelets improves in response to erythropoietin therapy. However these changes appear not to be related to the development of hypertension.

Platelet cytosolic free calcium concentration and parathyroid hormone: changing relationships with haemodialysis in end-stage renal disease.

1. Twelve patients receiving haemodialysis for end-stage renal failure were studied at a single dialysis session. Platelet cytosolic calcium concentration, plasma ionized calcium concentration and serum parathyroid hormone concentration were measured before dialysis, mid-dialysis and 30 min after dialysis. 2. Plasma ionized calcium concentration increased towards dialysate calcium concentrations, falling insignificantly after cessation of dialysis. Serum parathyroid hormone concentration fell by 39% during dialysis, with incomplete recovery afterwards. There was no overall change in platelet cytosolic calcium concentration. 3. Patients were divided into two subgroups: low parathyroid hormone (serum parathyroid hormone concentration less than 10 pmol/l) and high parathyroid hormone (serum parathyroid hormone concentration greater than 10 pmol/l). Before dialysis, values of platelet cytosolic calcium concentration or plasma ionized calcium concentration were not statistically different between the subgroups, but the platelet cytosolic calcium concentration was higher in the high-parathyroid hormone subgroup during and after dialysis. 4. Before haemodialysis there was a linear correlation between plasma ionized calcium concentration and platelet cytosolic calcium concentration, which disappeared during dialysis. In contrast, there was no relationship between serum parathyroid hormone concentration and platelet cytosolic calcium concentration before dialysis, but after dialysis a hyperbolic relationship was evident. 5. These results suggest that uraemic toxins may interfere with cytosolic calcium homoeostasis, allowing passive diffusion of extracellular calcium to influence the resting concentration, and that this effect is reversible by haemodialysis.