The rates of middle molecules in the blood in rats with different motor activity in alcoholic and non alcoholic exerimental hepatosis

Alcohol use disorders affect millions of individuals worldwide. The impact of these facts lies in the elevated social and economic costs. Liver metabolizes 75-98 % of ethanol that enters the organism. If the level of alcohol in the liver cells exceeds its degradation rate, alcoholic liver disease develops. The aim of the study was to determine the peculiarities of hepatotoxicity after simulation of of ethanol hepatosis and non-alcoholic steatohepatitis of high and low-motor active rats. Material and methods of investigation. The experiments were performed on on 72 white outbred male rats. The animals were divided into three groups: control, non-alkoholic hepatitis (NAH), ethanol hepatosis (EH). Each of group was subdivided – animals with high and low-motor activity (HA and LA). Contents of middle-mass molecules (MMM) were determined in the blood serum. Results. The analysis of the results shows that the levels of MMM in blood serum are increasing. Dystrophic changes that appear in a consequence of hypoxia are noted in NAH and EH. Both morphological and biochemical changes were more significant in HA animals. The degree of MMM accumulation depends on the severity of the pathology and motor activity of the animals. Less activity of MMM238 is observed in HA rats, which can be explained by the development of multi-organ pathology. Conclusions. During our investigations it was found the significant increasing of MMM levels in blood serum in rats with ethanol hepatosis and nonalkoholic hepatitis. The accumulation of MMM is not only a marker of endotoxication, they also increase the course of the pathological process, acquiring the roles of secondary toxins, affect the viability of all organs and systems. The degree of MMM accumulation depends on the moto activity of the animals and simulated pathology and is more significant in highly active animals, compared with low-active in the ethanol using. Less activity of MMM238 is observed in HA rats with EH, which can be explained by the development of multi-organ pathology, in particular, renal impairment. Morphological investigations showed that the grade of liver injury was more significant in HA rats.

Impact of Intradialytic Fiber Clotting on Dialyzer Extraction and Solute Removal: A Randomized Cross-Over Study

Previous studies revealed the importance of biocompatibility, anticoagulation strategy, and dialysis mode and duration on fiber blocking at the end of a hemodialysis session. The present study was set up in ten hemodialysis patients to relate fiber patency to dialyzer extraction and removal of small and middle molecules. With only 1/4th of the regular anticoagulation dose, and using a Solacea™ 19H and FX800 CorDiax dialyzer, fiber patency was quantified using 3D micro-CT scanning for different dialysis durations (i.e. 60, 120 and 240min). While Solacea™ showed good performance in all test sessions, fiber blocking in the FX800 CorDiax did not follow a linear process during dialysis, but was rather accelerated near the end of dialysis. Dialyzer extraction ratios were correlated with the percentages of open fibers. While the fiber blocking process affected extraction ratios (i.e. for phosphorus and myoglobin in the FX800 CorDiax), it had only minor impact on the removal of toxins up to at least 12kDa.

Interest of beta 2-microglubulin determination in chronic hemodialysis patients

The amylase beta 2-microglobulin (β2m) is a common complication of long-term hemodialysis, mainly responsible for osteo-articular manifestations including carpal tunnel syndrome (CTS). With the aim to study the various parameters involved in the increase of and determine the interest of dosage, we conducted a cross-sectional study of 104 chronic hemodialysis patients for more than 6 months. The mean age of patients was 52.82 years with a male predominance (62.5%). 61, 3% of our patients are anuric. They all have intermittent hemodialysis at 12 hours per week using a low permeability membrane in 52% of patients. The average age of dialysis layout is 42, 18 years. The mean duration of hemodialysis was 74 months. The mean serum β2m is 35, 24 mg / l. regarding complications, there were four confirmed cases of carpal tunnel syndrome and 12 deaths. Our study objectify the parameters involved in the increase of β2m: The current age, age advanced dialysis layout, lack of urine output, low permeability membrane and extended duration hemodialysis. Increased serum levels of β2m in our hemodialysis demonstrate the poor treatment of middle molecules. Its reduction can be made by improving the quality of dialysis and will avoid all the complications.

Highlights of the 12th Japan Bioanalysis Forum Symposium

Approximately 300 people associated with pharmaceutical industries, contractors, academic institutions and regulatory authorities attended the 12th Japan Bioanalysis Forum Symposium. The webinar was conducted from 9 to 11 March 2021. The theme of the symposium was ‘for the next generation’, and the event provided ‘an opportunity for young researchers in bioanalysis (including students)’ and ‘an opportunity to discuss new frontiers of bioanalysis’. The speakers focused on hot topics of bioanalysis, including biomarker analysis, patient centric sampling, virtual clinical trials, gene therapy, cancer genome medicine and therapeutic middle molecules. The symposium presented a platform for the discussion of the prospects and challenges facing bioanalysts working in the field of pharmacokinetics. This report presents the key issues discussed.

Pilot study to investigate differences in middle molecules, oxidative stress and markers of peripheral vascular disease in patients treated by high flux haemodialysis and haemodiafiltration

Background Dialysis patients have an increased risk of mortality. Recently treatment with haemodiafiltration (HDF) has been reported to reduce mortality, particularly cardiovascular mortality, compared to standard high-flux haemodialysis (HD). However, why HDF may offer a survival advantage remains to be determined. So, we conducted a pilot study to explore differences in middle-molecules, inflammation and markers of vascular disease in patients treated by HD and HDF. Methods Observational cross-sectional study measuring serum β2-microglobulin (β2M), Advanced Glycosylation End Products (AGEs) by skin autofluorescence (SAF), oxidative stress with ischaemia modified albumin ratio (IMAR) and peripheral vascular disease assessment using Ankle-Brachial Index (ABI), and arterial stiffness using Cardio-Ankle Vascular Index (CAVI). Results We studied 196 patients, mean age 69.1 ± 12.4 years, 172 (87.8%) treated by HD and 24 (12.2%) by HDF. Age, body mass index, co-morbidity and dialysis vintage were not different between HD and HDF groups. Middle molecules; β2M (31±9.9 vs 31.2±10 ug/mL) and SAF (2.99±0.72 vs 3.0±0.84 AU), ABI (1.06±0.05 vs 1.07±0.10) and CAVI (9.34±1.55 vs 9.35±1.23) were not different, but IMAR was higher in the HD patients (38.4±14.8 vs 31.3 ± 17.4, P = 0.035) Conclusions In this pilot observational study, we found patients treated by HDF had lower oxidative stress as measured by IMAR, with no differences in middle molecules. Lower oxidative stress would be expected to have diverse protective effects on the cardiovascular system Although we found no differences in ABI and CAVI, future studies are required to determine whether reduced oxidative stress translates into clinically relevant differences over time.

Flow Dynamic Analysis by Contrast-Enhanced Imaging Techniques of Medium Cutoff Membrane Hemodialyzer

<b><i>Introduction:</i></b> Medium cutoff (MCO) membranes represent an interesting innovation in the field of hemodialysis. Given the correlation between large (PM &#x3e;25 kDa) middle molecules (LMM) and clinical outcomes, the possibility to broaden the spectrum of solutes removed in hemodialysis with MCO membranes introduces a new perspective for end-stage kidney disease patients. Due to low diffusion coefficients of LMM, the use of convection is required to maximize extracorporeal clearance. High convective rates are achieved with high-flux membranes in hemodiafiltration, a technique not available in the US. In case of the MCO membrane, remarkable clearances of LMM are achieved combining the permeability of the membrane with a significant amount of internal convection. The mechanism of filtration-backfiltration inside the dialyzer enables effective removal of LMM in a technique called expanded hemodialysis (HDx). Given such theoretical explanation, it is important to demonstrate the blood and ultrafiltration rheology inside the MCO dialyzer. <b><i>Method:</i></b> This study for the first time describes flow dynamic parameters and internal cross-filtration, thanks to specific radiology and nuclear imaging techniques. <b><i>Results:</i></b> Flow dynamic analysis of the blood and dialysate compartment confirms excellent distribution of velocities and an excellent matching of blood and dialysate. Average blood flow velocity allows for wall shear rates adequate to avoid protein stagnation at the blood membrane interface and increase in blood viscosity. Cross-filtration analysis demonstrates a remarkable filtration/backfiltration flux reaching values &#x3e;30 mL/min at a blood flow of 300 mL/min and zero net filtration. <b><i>Conclusion:</i></b> The MCO dialyzer Theranova 400 appears to have a design optimized to perform expanded hemodialysis (HDx).

Study of oxygen compounds of acid character of Garbi Absheron field oil

The paper presents the results of fraction and structural-group composition of acids and phenols of Garbi Absheron field oil. Studied oil is heavy, tarry, low paraffinic. The acids and phenols have been purified from the oil with their differentiation on the free and associated (in the form of compound esters) forms. Free acids and phenols were purified from the oil via alkali extraction. Their associated forms were purified after saponification of the oil released from the samples. To study the structure-group content of acids and phenols, proton-magnetic resonance spectroscopy method was used. Obtained results showed that acidic oxygen-containing compounds in Garbi Absheron field oil predominantly consist of associated acids and phenols in the form of compound esters. Middle molecules of these phenols contain condensed naphtenic-aromatic cycles, the acids by their structure belong to the derivatives of hydrocarbons of paraffin series of CH3 residual-substitutes.

MO387CLINICAL PERFORMANCE, HEMOCOMPATIBILITY AND SAFETY OF A NEW DIALYZER WITH A MODIFIED POLYSULFONE MEMBRANE

Background and Aims Hemodialyzers containing membranes made from a blend of polysulfone and polyvinylpyrrolidone (PVP) are widely used. PVP makes the membrane material more hydrophilic to reduce interactions with plasma proteins and platelets. A modified spinning technique has been established to stabilize the PVP on the blood-side surface in the polysulfone dialysis membrane in the new dialyzer FX CorAL 600. The objective of the present study was to prove for this new dialyzer non-inferiority of performance in comparison to established dialyzers. Further, hemocompatibility and safety of the dialyzers were explored. Method In a multicenter, prospective, randomized, crossover study adult patients on online hemodiafiltration (HDF) were enrolled. They were treated for one week each with on-line HDF in post-dilution mode, and in randomized order with the dialyzers FX CorAL 600, FX 600, and FX CorDiax 600 (all Fresenius Medical Care, Bad Homburg, Germany). Blood samples were taken on the midweek session before start, at the end to analyze removal rate and at 60 min to determine clearance of ß2-microglobulin, myoglobin, urea, creatinine and phosphate. Further a pattern of hemocompatibility parameters and safety was evaluated. Assuming no carry over effect, linear mixed models were used for statistical analysis. Results The mean age of the 49 enrolled patients was 66.3±13.6 years, 76% were male. Treatments were performed in post-dilution mode with a mean blood flow &gt; 300 mL/min and a substitution volume &gt;19 L. The removal rate of ß2-microglobulin was 74.4, 70.4, and 73.1% for the FX CorAL 600, FX 600, and FX CorDiax 600 dialyzer, respectively. FX CorAL 600 proved to be statistically significantly non-inferior to FX 600 (p=0.0006) and to FX CorDiax 600 (p=0.036). The removal rate of FX CorAL 600 was by 4.0% (confidence interval 0.4 – 7.5%) significantly higher than with FX 600. The difference to FX CorDiax 600 was not significant. The clearance of ß2-microglobulin and myoglobin and the removal rate of myoglobin were significantly higher with the new dialyzer FX CorAL 600 than with the FX 600, and comparable to the FX CorDiax 600. Performance for small molecules was similar for all dialyzers. The complement factors C3a and C5a increased early in the treatment with a peak at 15 min, without differences between the three dialyzers for C3a, and for C5a with significantly lower increase at 15 min with FX CorAL 600 than for FX 600 (p=0.007); the difference of increase between FX CorAL 600 and FX CorDiax 600 was not significant (p=0.515). The course of sC5b9 was similar for all three dialyzers, with significantly lower increase at 15 min for both FX CorAL 600 (p=0.009) and FX CorDiax 600 (p=0.026) as compared to FX 600 and similar increase at 60 min for both FX CorAL 600 (p=0.573) and FX CorDiax 600 (p=0.386) as compared to FX 600. The area-under-the-curve for the course of sC5b-9 with FX CorAL 600 was significantly lower than with FX 600 (p=0.044) and comparable to FX CorDiax 600 (p=0.092). The leukocyte count showed a decrease in the first 15 min of the treatment, which recovered afterwards, similarly for all treatment phases with the different dialyzers. Further, the dialyzers did not differ with respect to adverse events. Conclusion All three dialyzers showed good performance, with higher removal rates for middle molecules with the new dialyzer FX CorAL 600 compared to the FX 600. Hemocompatibility profiles were mostly similar, with lower activation of C5a and of sC5b9 with FX CorAL 600 compared to FX 600. The new dialyzer provides comparable performance levels capable of delivering adequate treatment and good tolerability for the patient.

FC 134WATER SAVING COMPARISON AT DIFFERENT DIALYSATE FLOWS IN EXPANDED HEMODIALYSIS

Background and Aims Expanded hemodialysis (HDx) improves clearance of middle molecules as a target for uremia treatment. According to previously published results, high cut off filters have high enough clearance to allow diminishing of dialysate flow (Qd) without detrimental in HD adequacy outcomes with less water waste. According to World Health Organization (WHO), globally, almost 800 million people lack access to safe water and 2.5 billion lack access to optimal sanitation. Is our duty to search for ways to avoid water waste. Our aim is to describe the differences in water use between HDX patients receiving Qd 400 mL/min (Group 1) vs 500 ml/min (group 2) in HD patients from 10 renal clinics in Colombia as an alternative to reduce water waste in chronic HD as a strategy from Blue Planet Dialysis initiatives. Method We performed a Sub-analysis of CORHEX Study: We calculated water use at different Qd from our database: prospective, multicentre, observational cohort study of 992 adult patients undergoing chronic HD from 12 renal clinics in Colombia who were switched from high-flux HD to MCO therapy and observed for 12 months. All patients were prescribed with HDX three times a week for a minimum of 4 hours. We analysed patients with weight lower than 70 Kg at different Qd prescriptions to calculate water use at different Qd prescriptions and performed a prediction analysis, adjusting to Qd 400 mL/min, the whole potential population with weight lower than 70 Kg. Results 462 patients with weight lower than 70 Kg were analysed, 127 patients received Qd 400 mL/min. When diminishing Qd from 500 to 400 mL/min, 24 litres were saved per session per patient without detriment of middle molecule clearance. There were no differences in adequacy HD parameters between Qd prescriptions (Table 1 and 2). Conclusion HDX allows lowering of Qd to 400 mL/min without harm for patients and with remarkable savings of potable water: 24 Litres were saved in each session per patient that can be translated in almost 3500 Litres of water each year by patient which is enough for one year for 47 adults. (Based on the WHO minimum for basic health protection of at least 20 L per person/day) (2). We consider our results especially relevant since the World Health Organization estimates that within the next 5 years, over 50% of world population could reside in geographies lacking sufficient access to water. We hope that our results from the Blue Planet dialysis research group, alongside with Dr. Agar`s and Dr. Barraclough`s green nephrology initiatives, can help educate the nephrology community on the ecological impact of dialysis and can present an innovative solution to offer HD therapy even in countries suffering from limited access to potable water.

MO671COMPARASION OF EXPENDED HEMODIALYSIS USING A MEDIUM CUT-OFF DIALYZER VERSUS HEMODIAFILTRATION: AN OBSERVATIONAL PROSPECTIVE STUDY

Background and Aims Expended hemodialysis (HDx) with medium cut-off (MCO) membrane enables efficient depuration of middleweight uremic toxins, which play significant roles in inflammation and cardiovascular morbidity. Hemodiafiltration (HDF) is known for good removal of middle molecules but it requires more technical resources and well-functioning dialysis access. The aim of this study is to evaluate the efficacy of depuration of uremic toxins with a high-flux dialyzer during HDF session and with a MCO membrane (Theranova®) in HDx session and its impact on quality of life (QoL) in hemodialysis patients. Method In an open, single-centre, prospective observational clinical study, 28 adult stable HD patients without residual renal function were assigned to be treated by on-line HDF (HDF group) with the APS-21H dialyzer (polysulfone membrane, 2.1 m2, Asahi Kasei Medical Co., Japan) or by HDx (HDx group) with the Theranova® 400 (1.7 m2) and Theranova® 500 (2.0 m2) dialyzers (Baxter International Inc, USA). The study was conducted during 2019-2020 and completed after 12 months period. All patients were receiving maintenance high-flux membrane HDF treatment at least six months before they were enrolled in the study. Groups of patients were matched in age, sex, BMI, dialysis length and underlying disease. Complete blood count (CBC), renal function and inflammation, electrolytes, liver function tests, iron and nutritional status were evaluated at the beginning of the study and after 3, 6, 9 and 12 months. Pre and postdialysis levels for urea, creatinine, albumin, calcium, phosphorus, C-Reactive Protein, kappa and lambda free light chains (FLC), vitamin B12, β2 microglobulin levels were determined in each patient quarterly and reduction rate (RR) for uremic toxins were calculated. Furthermore single-pool Kt/V, dose of erythropoietin therapy (EPO) and vascular access were evaluated during the study, while bioimpedance analysis using Body composition monitor (Fresenius Medical Care, Germany) and QoL using SF-36 questionnaire (Kidney Disease Quality of Life Short Form-KDQOLTM-36) were evaluated at the end of observation period. The values have been reported as mean ±SD. Results There were 28 patients (14 in each group) mean age of 54.24 years (57.71±9.65 in HDx group vs 59.81±7.99 in HDF group). Median dialysis vintage was 4.77 years (5.33 in HDx group vs 6.46 in HDF group, p=0.55). Vascular access was native arteriovenous fistula in 23 patients, arteriovenous graft in 2 patients and tunnelled dialysis catheter in 3 patients (p=0.98). Kt/V was similar in both groups (1.57±0.31 vs 1.45±0.24, p=0.9), as well as weekly dose of EPO (4533.3±1922.3 vs 4233.3±1971.8, p=0.67). Patients in HDF group had a significantly higher interdialysis fluid overload (2,48±1,37 in HDx group vs 3,64±1,33 in HDF group, p=0.04), without difference in relation to the systolic and diastolic blood pressure values, as well as others BCM parameters. There were not significant differences in examined parameters of CBC, renal function and inflammation, electrolytes, liver function tests, iron and nutritional status at the beginning and at the end of the study. RR of small and middle molecules are presented in Table 1. Serum albumin level has decreased from 37.8 g/dL to 36.4 g/dL in 12 months during HDx treatment with maximal change of serum albumin level of -3.7% during that period (Figure 1). Evaluation of Kidney Disease Quality of Life Short Form at the end of study period in both groups is shown in Figure 2. Conclusion Compared to HDF, HDx with MCO membranes show greater RR for large middle molecules such as lambda FLC (45kD), while RRs for middle molecules- kappa FLC (23kD), β2 microglobulin (12kD) and small uremic toxins are similar. During one year of treatment with MCO membranes serum albumin levels remain stable. HDx treatment may improve quality of life, making an impact primarily in energy status and emotional satisfaction.