Blood Levels Research Articles

Baseline physiological data from anesthetized pigs in a VX intoxication model

Over the past fifty years, swine models have been used for organophosphorus intoxication studies. Among these studies and others on the swine model in general, some physiological data, especially cholinesterase activity highly impacted by organophosphorus compounds like nerve agent VX, still need to be completed. To support and compare our model to others, we have published the experimental protocol, the physiological values of 31 juvenile anesthetized pigs, and the 6h-follow-up of six supplementary anesthetized control animals and 7 VX-intoxicated pigs. We reported hemodynamics and respiratory parameters, blood levels in several biochemical parameters, blood gas, and complete blood count and compared them to the literature. We also focused on tissue and blood cholinesterase activities and detailed them for acetylcholinesterase and butyrylcholinesterase. After establishing a broad physiological data set consistent with the literature, we reported several cardio-respiratory parameters that seem more affected by an organophosphate intoxication, like heart rate, arterial blood pressure, cardiac output, and respiratory rate. Within the blood, oxygen saturation (SpO2), lactatemia, base excess, and glycemia can also be measured and associated with the other parameters to evaluate the life-threatening status.This swine model is currently used to develop and evaluate medical countermeasures against organophosphate nerve agent intoxications.

Polystyrene exposure induces lamb gastrointestinal injury, digestive disorders and inflammation, decreasing daily gain, and meat quality

Microplastics (MPs), recognized as an emerging environmental menace, have been extensively investigated in both marine and terrestrial fauna. This study is comprehensive to investigate how polystyrene (PS) affects ruminant animals. The experimental design comprised 24 individually housed lambs, divided into a CON group (diet without PS) and three PS-exposed (25 μm, 50 μm, 100 μm) groups, each with six lambs, the exposure of PS was 100 mg/day, and the duration of exposure was 60 days. The study yielded noteworthy results: (ⅰ) PS leads to a decrease in average daily gain along with an increase in feed conversion rate. (ⅱ) PS decreases rumen ammonia nitrogen. The rumen microbiota diversity remains consistent. However, the relative abundance of Bacteroidetes and Actinobacteria increased in the PS-exposed groups, while the relative abundance of Coriobacteriales_incertae_Sedis and Prevotellaceae_YAB2003_group decreased. (ⅲ) PS leads to decrease in hemoglobin, thrombocytocrit, and albumin levels in lamb blood, thus triggering oxidative stress accumulation, along with swelling of the kidneys and liver. (ⅳ) PS inflicts severe damage to jejunum, consequently impacting digestion and absorption. (ⅴ) PS reduces meat quality and the nutritional value. In conclusion, PS-exposure inhibited lambs’ digestive function, adversely affects blood and organs’ health status, reducing average daily gain and negatively influencing meat quality. PS particles of 50–100 μm bring worse damage to lambs. This research aims to fill the knowledge void concerning MPs’ influences on ruminant animals, with a specific focus on the meat quality of fattening lambs.

Curcumin affects autophagy of prolactinoma cells by upregulating miR-206 to exert antitumor effects.

We explored the effects of curcumin on the aberrant biological behaviors of prolactinoma cells and the downstream pathways through which curcumin exerts its antitumor effects. We used quantitative reverse transcription-polymerase chain reaction assays to measure miR-206 expression levels in peripheral blood samples from patients with prolactinoma before and after curcumin treatment. We also investigated the proliferation level, viability, and invasion ability of groups of cells treated with different concentrations of curcumin using 3-(4,5)-dimethylthiahiazo (-z-y1)-3-di-phenytetrazoliumromide (MTT) assays, cell cloning assays, and Transwell assays, respectively. Furthermore, we determined the levels of autophagy-related proteins and protein kinase B/mammalian target of the rapamycin (Akt/mTOR) signaling pathway-related proteins in each group of treated cells by western blot. Curcumin treatment upregulated miR-206 expression levels in the peripheral blood of patients with prolactinoma and in GH3 cells. Knockdown of miR-206 expression enhanced the proliferation and invasive ability of GH3 cells, while curcumin treatment effectively inhibited the aberrant biological behavior of GH3 cells enhanced by miR-206 knockdown. miR-206 knockdown also activated the Akt/mTOR signaling pathway and inhibited autophagy in GH3 cells, and these changes were effectively reversed by curcumin treatment. Thus, curcumin inhibited the Akt/mTOR signaling pathway and promoted cell autophagy by miR-206 upregulation, resulting in antitumor effects that inhibited prolactinoma cell proliferation and invasion.

A Retrospective Database Analysis to Investigate if Electrolytes in Venous Blood are Equivalent to the Levels in Arterial Blood.

In a critically ill patient, when an arterial blood sample is processed on an arterial blood gas (ABG) analyzer, it also measures electrolytes apart from analyzing the blood gases. The turnaround time for ABG analysis is way too less compared to the conventional electrolyte analysis with a serum sample. This study intends to investigate whether values of electrolytes estimated in arterial blood can substitute the routinely practiced method. This is a retrospective cross-sectional study. The source of data is patients' reports of serum electrolytes and ABG analysis from the Clinical Biochemistry laboratory, CIMS Teaching Hospital, Chamarajanagar between January and June 2021. The electrolytes report of 200 patients from whom both arterial and venous blood samples were sent to the Clinical Biochemistry laboratory on the same day and at the same time for analysis were selected. The data was compiled, compared, and correlated using a suitable statistical tool. The mean and standard deviation of sodium (135.62 ± 5.20 in venous vs 134.08 ± 8.49 in arterial blood), potassium (4.20 ± 0.64 vs 3.80 ± 0.75), and chloride (102.28 ± 4.99 vs 96.33 ± 8.11) were observed. However, when the concordance correlation coefficient and Bland-Altman plot analysis were made there was no agreement between electrolytes analyzed on serum in an autoanalyzer with that of ABG analyzer. We conclude that the electrolytes measured by a conventional autoanalyzer on a serum sample cannot be replaced by values analyzed on a blood gas analyzer. Devaki RN, Kasargod P, Roopa Urs AN, Chandrika N. A Retrospective Database Analysis to Investigate if Electrolytes in Venous Blood are Equivalent to the Levels in Arterial Blood. Indian J Crit Care Med 2024;28(5):442-446.

Biological profile of secondary hyperparathyroidism in chronic renal failure patients

Secondary hyperparathyroidism (SHPT) is a common complication of chronic kidney disease, especially in hemodialysis patients. It is characterized by overproduction of parathyroid hormone in response to chronic deterioration of renal function. The aim of this retrospective study was to determine the average time to develop secondary hyperparathyroidism, analyze the initial phosphocalcic profiles, and evaluate the evolution of these profiles in hemodialysis patients, following the recommendations of the KDOQI 2003 and KDIGO 2009 groups. The study included 134 hemodialysis patients from two private dialysis centers. Data were collected from patients' medical records and analyzed using SPSS software. The results showed that the average time to develop secondary hyperparathyroidism was 3.15 years according to KDOQI recommendations and 4.64 years according to KDIGO recommendations based on parathyroid hormone levels. The initial assessment revealed an imbalance in phosphocalcic metabolism in approximately 20.1% to 22.5% of patients, a proportion that increased to 27.5% during follow-up, according to both sets of recommendations. In conclusion, secondary hyperparathyroidism is an inevitable complication of chronic kidney disease, affecting the vital and functional prognosis of hemodialysis patients. Diagnosis primarily relies on parathyroid hormone levels, associated with phosphocalcemia, vitamin D, and PAL analyses. Treatment aims mainly to maintain blood levels of calcium, phosphorus, and vitamin D within recommended limits and to reduce parathyroid response to phosphocalcic disturbances.

PACAP38/mast-cell-specific receptor axis mediates repetitive stress-induced headache in mice.

Pain, an evolutionarily conserved warning system, lets us recognize threats and motivates us to adapt to those threats. Headache pain from migraine affects approximately 15% of the global population. However, the identity of any putative threat that migraine or headache warns us to avoid is unknown because migraine pathogenesis is poorly understood. Here, we show that a stress-induced increase in pituitary adenylate cyclase-activating polypeptide-38 (PACAP38), known as an initiator of allosteric load inducing unbalanced homeostasis, causes headache-like behaviour in male mice via mas-related G protein-coupled receptor B2 (MrgprB2) in mast cells. The repetitive stress model and dural injection of PACAP38 were performed to induce headache behaviours. We assessed headache behaviours using the facial von Frey test and the grimace scale in wild-type and MrgprB2-deficient mice. We further examined the activities of trigeminal ganglion neurons using in vivo Pirt-GCaMP Ca2+ imaging of intact trigeminal ganglion (TG). Repetitive stress and dural injection of PACAP38 induced MrgprB2-dependent headache behaviours. Blood levels of PACAP38 were increased after repetitive stress. PACAP38/MrgprB2-induced mast cell degranulation sensitizes the trigeminovascular system in dura mater. Moreover, using in vivo intact TG Pirt-GCaMP Ca2+ imaging, we show that stress or/and elevation of PACAP38 sensitized the TG neurons via MrgprB2. MrgprB2-deficient mice showed no sensitization of TG neurons or mast cell activation. We found that repetitive stress and dural injection of PACAP38 induced headache behaviour through TNF-a and TRPV1 pathways. Our findings highlight the PACAP38-MrgprB2 pathway as a new target for the treatment of stress-related migraine headache. Furthermore, our results pertaining to stress interoception via the MrgprB2/PACAP38 axis suggests that migraine headache warns us of stress-induced homeostatic imbalance.

The role of tryptophan and its derivatives in musculoskeletal pains: A systematic review and meta-analysis.

Studies present ambiguous findings regarding the role of tryptophan and its metabolites, kynurenine and serotonin in chronic musculoskeletal pain. This systematic review aimed to investigate the expression of tryptophan and its metabolites, serotonin and kynurenine in patients with local and generalized chronic musculoskeletal pain in comparison with pain-free controls. An electronic search was conducted in the databases MEDLINE, CINAHL, EMBASE, the Cochrane Central Registry of Controlled Trials (CENTRAL) and Web of Science for clinical and observational trials from the beginning of each database to 21 April 2023. Out of 6734 articles, a total of 17 studies were included; 12 studies were used in the meta-analysis of serotonin, 3 regarding tryptophan and 2 studies for a narrative synthesis regarding kynurenine. Risk of bias was assessed using the quality assessment tool for observational cohort and cross-sectional studies of the National Heart, Lung, and Blood Institute, while the certainty of evidence was by GRADE. All included studies showed a low risk of bias. The meta-analysis showed lower blood levels of tryptophan (p < .001; very low quality of evidence) and higher blood levels of serotonin (p < .001; very low-quality evidence) in patients with generalized musculoskeletal pain, when compared to pain-free individuals. In local chronic musculoskeletal pain, there were higher blood levels of serotonin (p=.251; very low quality of evidence) compared to pain-free individuals. Regarding kynurenine, the studies reported both higher and lower blood levels in generalized chronic musculoskeletal pain compared to pain-free individuals. The blood levels of tryptophan and its metabolites serotonin and kynurenine seem to influence chronic musculoskeletal pain.

Serum ferritin levels in children of simple febrile seizures in tertiary institute

Background: Seizure onset is impacted by a number of variables, including genetics, diet, geography, concurrent illnesses, metabolic state, history of head trauma, and blood levels of particular minerals. Due to the fact that iron is required for the proper functioning of a number of enzymes and neurotransmitters in the central nervous system, low blood ferritin levels may lower the seizure threshold. The aim of the study was to determine the association between serum ferritin levels and simple febrile seizures. Methods: The present observational study was conducted in department of pediatrics of a tertiary institute and included a total of 200 study subjects who visited to pediatric OPD/emergency room with febrile illness. The data was collected with the help of a structured clinical proforma. The collected data was recorded in Microsoft Excel sheet and statistical analysis was done with the help of SPSS version 21.0. Results: In our study, the maximum number of the study subjects were between 13 and 24 months (35%) with male to female ratio 1.2:1. In patients with simple febrile seizure the mean Hb level was 9.1±1.4 gm/dl, the mean MCV was 72.6±7.4 f1, mean MCH was 23.73±3.2 pg and mean serum ferritin was 13.4±9.5 ng/ml as compared to those patients with febrile illness without seizure the mean Hb level was 11.5±1.2 gm/dl, mean MCV was 82.1±5.5 f1, mean MCH was 29.6±2.9 pg and mean serum ferritin was 33.6±20.2 ng/ml. Conclusions: It is concluded that the low serum ferritin level is a risk factor of simple febrile seizure.

Acute kidney injury in lamotrigine-induced DRESS syndrome.

We present a case of lamotrigine-triggered DRESS (drug reaction with eosinophilia and systemic symptoms) syndrome with acute kidney injury stage 3. A 17-year-old girl with known epilepsy treated with lamotrigine presented with acute kidney injury as well as skin eruption, fever, and apathy. Extended diagnostics, considering infectious and autoimmune diseases, remained unremarkable. Lamotrigine blood levels were within the target range. Kidney biopsy showed acute interstitial nephritis with tubular necrosis. Methylprednisolone pulse therapy led to an improvement in kidney function; skin eruption and neurological symptoms resolved. During the hospital stay, the girl admitted to inconsistent and variable intake of lamotrigine, occasionally resulting in notable overdosing. This report demonstrates that acute kidney injury in lamotrigine-induced DRESS syndrome is an acute interstitial nephritis with tubular necrosis, an aspect that has not been deeply characterized so far. Additionally, we aim to elevate awareness towards non-adherence as cause of disease, especially among the adolescent population.

Ingestion of β-nicotinamide mononucleotide increased blood NAD levels, maintained walking speed, and improved sleep quality in older adults in a double-blind randomized, placebo-controlled study.

The study evaluated how ingestion of nicotinamide mononucleotide (NMN) for 12weeks by older adults affected blood nicotinamide adenine dinucleotide (NAD +) levels and physical function, particularly walking function. Information concerning sleep, and stress was also collected as secondary endpoints. In this randomized, placebo-controlled, double-blind, parallel-group comparison study, 60 participants were randomly allocated into a placebo group or NMN group. Members of the NMN group consumed 250mg/day NMN for 12weeks. Motor function tests, blood NAD metabolite analysis, and questionnaires were conducted at the start of the study and 4 and 12weeks after intake. This trial was registered at umin.ac.jp/ctr as UMIN000047871 on June 22nd, 2022.At primary outcome, at both 4weeks and 12weeks, the NMN and placebo groups had no significant differences in a stepping test. At secondary outcomes, after 12weeks of NMN intake, the NMN group had a significantly shorter 4-m walking time than the placebo group as well as significantly higher blood levels of NAD + and its metabolites. A significant negative correlation was observed between the change in the 4-m walking time and the change in blood NAD + , N1-methyl-2-pridone-5-carboxamide (2-PY), and N1-methyl-4-pridone-3-carboxamide (4-PY) at 12weeks. The NMN group had improved sleep quality at 12weeks relative to the placebo group as evidenced by lower scores for "Daytime dysfunction" and "Global PSQI" on the Pittsburgh Sleep Questionnaire. No adverse effects related to test substance consumption were observed. Together, these results indicate that NMN intake could increase blood NAD + levels, maintain walking speed, and improve sleep quality in older adults. Interventions involving NMN aimed at maintaining walking speed could contribute to extended healthy life expectancy.

Treatment-resistant hiccups during general anesthesia possibly caused by remimazolam: a case report

BackgroundPrevious reports have described hiccups during general anesthesia that were possibly induced by drugs, including benzodiazepines. However, there are few reports of hiccups caused by remimazolam.Case presentationA 75-year-old woman underwent corneal transplantation under general anesthesia with remimazolam. She presented with hiccups once the effects of muscle relaxants used during induction wore off, which persisted even after various treatments, such as the administration of antipsychotic drugs. However, when remimazolam administration was terminated after surgery to awaken the patient, the hiccups stopped and did not recur after extubation. Evaluation of predicted blood levels of remimazolam suggests that higher levels of remimazolam might cause hiccups.ConclusionRemimazolam might induce hiccups during general anesthesia. Anesthesiologists should consider administering muscle relaxants or changing the anesthetic in cases of refractory hiccups under general anesthesia.

Correlation of drug dose estimated from national prescription registers with mean blood level of antiseizure medication in pregnancy.

The purpose was to examine the correlation of antiseizure medication drug dose estimated from prescription fill records from prescription registers with blood levels during pregnancy. We conducted a Nation-wide study of mothers who gave birth in Denmark between 1 January 2014 and 31 December 2018 using data from Danish Prescription and Laboratory Registers. We identified mothers with blood level measurements of antiseizure medication. The main exposure was estimated antiseizure medication dosage estimated from pregnancy-filled prescriptions in the Danish Prescription Register. The main outcome was the correlation of estimated dose with mean blood level of antiseizure medication in pregnancy. For privacy reasons, the number of blood level measurement and prescription fills were rounded to nearest 10, but proportions reported as exact values. Among 298 560 pregnancies, we identified pregnancies with recorded prescription fill from the prescription register for valproate (N = 90), lamotrigine (N = 1360), levetiracetam (N = 340), topiramate (N = 100), and carbamazepine (N = 60). In these pregnancies, blood level measurements were available in 50 (53%) pregnancies for valproate, 850 (62%) pregnancies for lamotrigine, 320 (93%) pregnancies for levetiracetam, 50 (68%) pregnancies for carbamazepine, and 40 (35%) pregnancies for topiramate. Pearsons's correlation coefficients for the correlation of estimated antiseizure medication dose with mean blood levels were 0.67 (p < 0.0001) for valproate, 0.63 (p < 0.0001) for lamotrigine, 0.63 (p < 0.0001) for levetiracetam, 0.76 (<0.0001) for carbamazepine and 0.89 (<0.0001) for topiramate. Dose of antiseizure medication estimated from prescription fills was a good proxy for blood levels and thus for biological exposure in pregnancy, suggesting that administrative prescription fill records may be a valuable resource for estimating exposure to antiseizure medication in pregnancy.

#1431 Risk factors of adverse cardiovascular events in the early postoperative period in kidney transplant recipients

Abstract Background and Aims Kidney transplantation improves quality of life and increases life expectancy in patients with Stage 5 Chronic Kidney Disease. However, the frequency of adverse cardiovascular events in the postoperative period remains high and reduces the effectiveness of transplantation. The aim was to study the incidence of cardiovascular risk factors in the preoperative period in kidney transplant recipients with adverse cardiovascular events in the early postoperative period. Method The study included 51 kidney transplant recipients whose age was 46.1 ± 4.6 years. 12 (23.5%) recipients had adverse cardiovascular events (acute myocardial infarction, unstable angina, rhythm disturbance, complicated hypertensive crisis) in the early postoperative period. 39 (76.5%) recipients with uncomplicated postoperative period formed the comparison group. The study groups were comparable by gender and age. Results Traditional cardiovascular risk factors were assessed. The study groups had comparable rates of arterial hypertension, obesity, hyperglycemia, dyslipidemia and smoking in the preoperative period and graft function in the postoperative period. The study groups didn't have significant differences in the incidence of anemia and hemoglobin blood concentration. But the erythropoietin blood level in the preoperative period in recipients with adverse cardiovascular events was higher—38.5 (23.5-155.0) mU/mL versus 16.0 (10.0-32.0) mU/mL, p &amp;lt; 0.01, and hypererythropoietinemia was diagnosed more often—58.3% (n = 7) versus 25.6% (n = 10), p &amp;lt; 0.05, than in patients without cardiovascular events. The incidence of elevated interleukin-6 blood level in the preoperative period was higher in recipients with adverse cardiovascular events—25.0% (n = 3) versus 5.1% (n = 2), p &amp;lt; 0.05. At the same time C-reactive protein levels and white blood cell counts were not significantly different. Recipients with adverse cardiovascular events had a lower left ventricular ejection fraction—59.5 ± 10.6% versus 65.6 ± 7.1%, p &amp;lt; 0.05, and a higher incidence of left ventricular diastolic dysfunction in the preoperative period—75.0% (n = 9) versus 35.9% (n = 14), p &amp;lt; 0.05, than in recipients without cardiovascular complications. Conclusion Kidney transplant recipients with adverse cardiovascular events in the early postoperative period had a higher incidence of left ventricular diastolic dysfunction, increased blood levels of erythropoietin and interleukin-6 and a lower left ventricular ejection fraction before transplantation than recipients with an uncomplicated postoperative period.

#2343 Fabry disease screening in patients with chronic kidney disease and dialysis at a single medical center in Taiwan

Abstract Background and Aims Chronic Kidney Disease (CKD) poses a significant global health challenge, with diverse contributing factors impacting its progression. Among these, Fabry Disease, a rare lysosomal storage disorder, has emerged as a potential complicating factor. Despite its relevance, the prevalence of Fabry Disease among CKD patients, particularly in Taiwan, remains insufficiently explored. Recognizing and understanding the occurrence of Fabry Disease in CKD populations are crucial for comprehensive patient care and may have implications for treatment strategies. Additionally, investigating the relationship between kidney function and Plasma α-Gal A activity is essential. α-Gal A, the enzyme deficient in Fabry Disease, may experience altered activity levels in the presence of compromised kidney function. Clarifying this connection is pivotal for gaining insights into the intricate interplay between Fabry Disease and CKD. This study aims to achieve three key objectives (1) Determine Fabry Disease Prevalence among CKD Patients in Taiwan.(2) Investigate the Impact of Kidney Function on Plasma α-Gal A Activity.(3) Characterize Gene Mutation Patterns and Demographic Data of Fabry Patients in CKD Population in Taiwan: Method Fabry Disease assessment involved a two-stage process, tailored to the patient's gender. Males underwent a dry blood test measuring α-galactosidase activity; samples below 15% of the adult average were deemed positive. Females were screened based on lyso-Gb3 levels in whole blood; a result exceeding 0.8 ng/ml was considered positive. Positive screenings prompted genomic DNA extraction, and the GLA gene's seven exons and intron 4 segment ([GRCh38/Hg38] chrX:101,399,610-101,399,874) were PCR-amplified. Sanger sequencing identified pathogenic variants. This gender-specific, multi-step approach ensures precision. Defined thresholds and advanced genomic techniques enhance diagnostic accuracy. The method provides a robust means of identifying Fabry Disease, supporting early intervention and personalized treatment. Results Study Recruitment: Initially, 2000 subjects enrolled, but 155 withdrew, yielding a final cohort of 1845 patients for analysis. Screening and Referral: Among 1602 subjects, 26 males (1.6%) were referred for DBS LysoGB3 check-up due to low serum α-Gal A activity. In a cohort of 243 females, 6 individuals (2.4%) underwent GLA genetic sequencing due to elevated lyso-GB3 levels. Clinical Presentation - Male Case: A male with acute stroke symptoms and CKD stage 3b (eGFR 34.5 mL/min/1.73 m²) showed significantly low DBS α-Gal A activity (0.48 μmol/hr) and elevated Plasma LysoGB3 (28.08 ng/ml).GLA genetic sequencing revealed a specific deletion (c.1072-1074delGAG, p. (Glu358 del)) in exon 7, leading to a Fabry disease diagnosis, classifying it as a specific type. The family pedigrees of this male patient are visually represented in Fig. 1, illustrating the genetic relationships within the family. Clinical Presentation - Female Case: A female on hemodialysis for four years, cause unknown, exhibited a DBS LysoGB3 level of 0.5 ng/ml. GLA genetic sequencing identified a nucleotide variation (c.640-801G&amp;gt;A) in intron 4, establishing a diagnosis of Fabry disease with the cardiac variant (301500). Fig. 2 demonstrates the relationship between serum DBS GLA activity and different stages of CKD. Our analysis reveals that in CKD stages 1, 2, 3 (3a and 3b), and 4, the serum GLA activity is significantly higher compared to stage 5. Conclusion The prevalence of Fabry Disease in our cohort was 0.1% (2 out of 1845 individuals), featuring one female with the cardiac variant and one male with the class type. Our investigation unveiled a significant rise in serum GLA activity in CKD stages 1–4 compared to stage 5, suggesting potential implications for early CKD detection. Moreover, serum α-Gal A activity displayed a notable correlation with age, BUN, and eGFR across CKD stages 1–5, questioning its status as a biomarker for advanced CKD and aging in CKD patients. This prompts consideration for a large-scale study to validate this hypothesis.

#1502 Short-term effects of gadolinium and gadoteric acid on kidney — impact on inflammation

Abstract Background and Aims The nephrotoxicity of free gadolinium [Gd (III)] has been reported, raising concerns about the safety of gadolinium-based contrast agents (GBCA), widely used in magnetic resonance imaging. The GBCA with macrocyclic structure, such as gadoteric acid (Gd-DOTA), appear as more stable. Using human proximal tubular cells cultures, we found that free Gd (III) triggers death by apoptosis and necrosis, and increases the expression of modulators of inflammation, hypoxia and fibrosis, which are known as major risk factors for the development and worsening of kidney disease. Our aim was, using animal models, to evaluate the short-term effects of Gd (III) and of gadoteric acid on inflammatory biomarkers, at blood and renal tissue levels. Method Wistar rats were exposed to a single dose of Gd (III) or Gd-DOTA (groups A and B, respectively); a control group was also included (C). After 48h, blood and kidney samples were collected. The plasma levels of interleukin (IL)6 and transforming growth factor (TGF) beta 1 were assessed by ELISA; the kidney gene expression of IL6 (il6), TGF beta 1 (tgbf1) and nuclear factor-kappa B (nfkb1) was determined through qPCR. Results Gd (III) group (A) presented higher circulating IL6 levels (P = 0.001) and increased kidney gene expression of il6 (P = 0.013), tgbf1 (P = 0.006) and nfkb1 (P = 0.011) than the control group (C); compared to Gd-DOTA group (B), the IL6 plasma concentration (P = 0.005) and tgbf1 mRNA levels were also higher (P = 0.004). The Gd-DOTA group (B) presented increased gene expression of il6 (P = 0.006) than the control group (C). Conclusion Short-term exposition to free Gd (III) induced an inflammatory response, shown by the increase in circulating IL6, and in kidney gene expression of il6, tgbf1 and nfkb1. The exposition to Gd-DOTA was associated with a lower inflammatory response, showing only increased kidney gene expression of il6. Despite the much safer profile for Gd-DOTA, further studies are warranted, evaluating other biomarkers, to clarify the short-term, and even the long-term effects, of these compounds. Acknowledgements This work was financed by FCT through the project 2022.08400.PTDC; FCT, in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences—UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy—i4HB.

#2751 CD14++CD16+ monocytes and CD8+ T cells are independently associated with annual change in eGFR and proteinuria in kidney transplant recipients

Abstract Background and Aims Immune cellular responses are implicated in all clinical aspects of kidney transplantation. There is little data regarding the additional value of regular monitoring of immune cells subsets expression in kidney transplant recipients (KTRs). The aim of our prospective study was a longitudinal follow-up analysis of monocytes subpopulations, natural killer (NK) cells and lymphocytes subsets including regulatory T cells (Tregs) in the circulation of KTRs and potential clinical correlations. Method 48 stable KTRs were initially enrolled. All KTRs were under immunosuppressive regimen with corticosteroids, mycophenolate mofetil or mycophenolate acid and calcineurin inhibitors (CNIs). Exclusion criteria were history or development during follow-up of acute rejection, cardiovascular disease, malignancy and active or chronic infections. Patients were followed for 12 months. The peripheral blood immune cell subsets CD14++CD16-, CD14++CD16+ and CD14+CD16++ absolute values and percentages out of total monocytes and NK cells (CD3+CD16+56+), CD3-CD19+ B lymphocytes, CD3+ CD4+ T cells, CD3+CD8+ T cells and Tregs (CD4+CD25+ FoxP3+) absolute values and percentages out of total lymphocytes were measured by flow cytometry at baseline (T0) and after 12 months (T1). Clinical and laboratory parameters were recorded at T0 and T1. Delta (Δ) eGFR and Δ spot urine protein to creatinine ratio (ΔUPCR) were defined as their respective differences between T1 and T0. Likewise, Delta (Δ) of immune cells subtypes or laboratory indices was defined as their respective difference between T1 and T0. Results 35 KTRs (mean age 53 ± 9.28 years, 71% males, mean transplant vintage 96 ± 66 months, 63% on Tacrolimus and 37% on Cyclosporine) were included in the final analysis. Significant differences were observed between T0 and T1 in monocytes number (653 ± 244 and 538 ± 197/μL respectively, p = 0.001), monocytes percentage (7.6 ± 2.9 and 6.6 ± 2.2% respectively, p = 0.006) as well as in the number of classical CD14++CD16- monocytes (534 ± 225 and 452 ± 185/μL respectively, p = 0.04). The rest immune cells subsets did not show any significant differences between T0 and T1. Mean eGFR declined from 58 ± 17 at T0 to 53 ± 18 ml/min/1.73 m2 at T1 (p = 0.004). No significant changes were observed between T0 and T1 in median UPCR [0.16 (IRQ, 0.09-0.56) at T0 and 0.16 (IQR, 0.10-0.70) gr protein/gr creatine at T1, p = 0.489], in median CRP [4.0 (IQR, 3-7) at T0 and 5 (IQR, 2.5-7.5) mg/L) at T1, p = 0.919], in mean ESR (20 ± 14 at T0 and 22 ± 19 mm/hour at T1, p = 0.381) or other parameters, including CNIs blood levels. ΔeGFR was correlated with the T0 percentage of monocytes (rho = 0.359, p = 0.037), the T0 number and T0 percentage of CD14++CD16+ monocytes (rho= 0.502, p = 0.003 and rho = 0.438, p = 0.008 respectively). On the other hand, a borderline inverse correlation was observed between ΔeGFR and ΔCD14++CD16+ monocytes number (rho = −0.339, p = 0.05). Additional correlates of ΔeGFR included serum albumin at T0 (rho = 0.395, p = 0.021) and Δuric acid (rho = 0.567, p = 0.000). At stepwise linear regression analysis, CD14++CD16+ monocytes (β = 0.338, p = 0.04) and Δuric acid (β = 0.477, p = 0.006) remained independent significant correlates of ΔeGFR. ΔUPCR was significantly correlated with the percentage of B-lymphocytes (rho = 0.385, p = 0.027) and CD4+ T-cells (rho = 0.352, p = 0.044) at T0, and inversely correlated with the T0 percentage of T-lymphocytes (rho = −0.402, p = 0.02) and CD8+ T cells (rho = −0.603, p = 0.000) as well as with Δhemoglobin (rho = −0.385, p = 0.027). At stepwise linear regression analysis, only the CD8+ T cells percentage at T0 remained independently correlated to ΔUPCR (β = –0.379, p = 0.03). Conclusion The results of our study indicate that intermediate CD14++CD16+ monocytes are associated with annual eGFR change whereas CD8+ T cells negatively correlate with the annual trend of proteinuria change in KTRs. Further research is needed to clarify underlying pathophysiological mechanisms for these findings. Finally, future studies including larger cohorts and with longer follow-up are required to specify the potential utility of immune subpopulations monitoring as potential prognostic biomarkers in KTRs.

#2881 Glucose free-based solutions reduce oxidative stress in peritoneal dialysis: support for a new promising approach

Abstract Background and Aims Oxidative stress (OxSt) and inflammation are common in CKD which are known CV and mortality risk factors. In peritoneal dialysis (PD) OxSt and Inflammation even increase due to the use of glucose-based solutions. This study evaluated in PD patients the effect of 3 and 6 months treatment with icodextrin-based glucose-free solutions on OxSt and inflammation. Method In 15 PD patients using a molecular biology approach, p22phox protein expression (Western blot), subunit of NADPH oxidase, essential for OxSt activation, MYPT-1 phosphorylation state, marker of RhoA/Rho kinase pathway activity, deeply involved in OxSt generation (Western blot) and Malondialdehyde (MDA) production (fluorimetric assay), marker of lipid peroxidation, have been evaluated. High-sensitive C Reactive Protein (hsCRP) (immunoturbidimetric assay) and interleukin (IL)-6 (chemiluminescence assay), have been used as marker of inflammation. Results p22phox protein expression, MYPT-1 phosphorylation, and MDA level were reduced after 3 months from the start of icodextrin based solutions (1.28 ± 0.18 d.u. vs 1.50 ± 0.19, p = 0.049; 0.89 ± 0.03 vs 0.98 ± 0.03, p = 0.004; 4.20 ± 0.18 nmol/mL vs 4.84 ± 0.32 nmol/mL, p = 0.045, respectively). In a subgroup of 9 patients who continued the treatment up to 6 months MYPT-1 phosphorylation was further reduced at 6 months compared to baseline (0.84 ± 0.06 vs 0.99 ± 0.04, p = 0.043), while p22phox protein expression was reduced only at 6 months from baseline (1.03 ± 0.05 vs 1.68 ± 0.22, p = 0.021). In this subgroup MDA blood level was reduced at 6 months from baseline (4.03 ± 0.24 nmol/mL vs 4.68 ± 0.32, p = 0.024) and was also significantly reduced vs 3 months (4.03 ± 0.24 vs 4.35 ± 0.21, p = 0.008). IL-6 level, although reduced compared to baseline, did not reach statistical significance both at 3 and 6 months, while hsPCR was unchanged. Conclusion The icodextrin-based PD solutions mediated reduction of OxSt related proteins and inflammation we have provided, although obtained in a small cohort of patients and for a limited time duration, strongly support the rationale of using in PD osmo-metabolic agents-based solutions replacing the currently used glucose-based fluids. The results of ongoing studies with these agents in PD patients will provide definitive proofs to better preserve peritoneal membrane integrity and residual renal function but also to reduce CV risk factors such as OxSt and inflammation.

#2733 Osteoporosis and CKD-MBD in hemodialysed patients: efficacy and safety of denosumab

Abstract Background and Aims Osteoporosis is a chronic progressive disease characterized by low bone mass and deterioration of bone tissue which results in a rapid increase in fracture risk. Osteoporosis in hemodialysed patients with Chronic Kidney Disease – Mineral and Bone Disorder (CKD-MBD) is a debilitating clinical condition with complex therapeutic management. In fact, most of the drugs commonly used to counteract osteoporosis are generally contraindicated when Glomerular Filtration Rate (GFR) is lower than 35 ml/min. Denosumab is approved for the treatment of osteoporosis and does not require dose adjustment in case of impaired renal function. For this reason, since 2014 this drug has been used by nephrologists for the treatment of osteoporotic patients also in hemodialysis. Although the interest and diffusion in this specialized clinical field are growing, only few data are still available in literature. The aim of the study was to evaluate the efficacy and safety of Denosumab in long-term therapies (up to 68 months of therapy) in hemodialysed patients. Method Since December 2013, a total of 22 hemodialysed patients have been treated with Denosumab annually (4) or semi-annually (18) for up to 68 consecutive months. During the treatment, we monitored the blood levels of calcium, phosphorus, PTH, beta-CrossLaps and bone alkaline phosphatase (BALP), T-SCORE and fracture risk (FR) using Qualitative UltraSonography (QUS) or Computerized Bone Mineralometry (CBM) every 30 months and every 2 years, respectively. Results 4/22 patients completed at least 48 months of follow-up, observing a substantial stability of the mean values of blood calcium (from 9.09 mg/dl to 8.63 mg/dl) and phosphorus (from 5.2 mg/dl to 4.1 mg/dl), against a reduction in the mean values of B-CrossLaps and bone alkaline phosphatase (BALP) (respectively from 2975.32 to 1452.75 pg/ml and from 32.09 to 14.58 mcg/L). Comparison of QUS exams demonstrated improved T-score (mean values from −4.71 to −4.17) and reduced fracture risk (mean value from 13% to 8%). The results of the CBMs confirmed the improvement of the bone disease at the lumbar spine (T-score from −3.8 to 2.8 after 2 years of treatment) and the substantial stability of the T-score at the femoral level (from −3.5 to −3.2 after 2 years of treatment). It should be noted that in our samples two patients achieved the longest observation period (68-month follow-up in continuous therapy), in which we observed improvement of the bone disease also at the femoral level compared to the start of treatment (T-score from −3.9 to −2.8 after 48 months of treatment). Only one fracture has been recorded. Conclusion The results obtained confirm efficacy and safety of Denosumab in reducing the risk of fractures, with the improvement of BMD and T-score at the lumbar spine using Computerized Bone Mineralometry (CBM). We underline that close biochemistry monitoring and careful evaluation of the therapeutic procedures before and after Denosumab administration helped us to minimise and promptly correct adverse effects due to hypocalcemia. In addition we are particularly satisfied to report in most patients a significant reduction of pain and an improvement of mobility. The results collected are consistent with acceptable safety and demonstrated efficacy of Denosumab in hemodialysis patients.

#2137 Systemic inflammation as a main factor in CKD-MBD development

Abstract Background and Aims Chronic kidney disease (CKD) is a progressive disorder that affects more than 10% of the general population worldwide. Despite great achievements in the field of treatment of CKD, patient mortality remains at a high level. Research into the pathophysiology and correction of this condition in the early stages of development is ongoing. The aim of the study was to establish relationship between systemic inflammation and mineral disorders in any stage of CKD. Method We investigated 5 groups of patients with different stages (I to V) of CKD 19-22 persons in each (total amount 106 patients), aged 49,6 years and 19 healthy individuals aged 41,9 years as control group. We examined the blood level of interleukin 1 beta (IL-1β), interleukin 6 (IL-6), interleukin 8 (IL-8), C-reactive protein (CRP) and c-terminal fragment of fibroblast factor 23 (FGF-23) in all patients and control group. Results The course of CKD is accompanied by systemic inflammation, which is characterized by an increase in IL-1β in the I stage of chronic kidney disease ((13,41 ± 3,36) pg/ml) (p &amp;lt; 0.01), IL-6 in the III stage of chronic kidney disease (29,31 ± 5,49) pg/ml (p &amp;lt; 0.01), IL-8 in the II stage of chronic kidney disease ((11,56 ± 2,72) pg/ml) (p &amp;lt; 0.05) and CRP in the IV stage of chronic kidney disease ((7,22 (1,81; 10,8)) mg/ml (p &amp;lt; 0.01), compared to the control group ((5,93 ± 3,65), (6,01 ± 1,67), (7,99 ± 1,17) pg/ml and (0,66 (0,41; 0,85)) mg/ml accordingly), and progresses with a decrease in glomerular filtration rate. Growing already at the initial stage of chronic kidney disease, the level of FGF-23 reached a statistical difference from the control group at the 2nd stage of CKD. A strong statistically significant relationship was found between FGF-23 and IL-6 (R = 0.84; p &amp;lt; 0.01), IL-1β at GFR &amp;gt; 15 ml/(min•1.73 m2) (R = 0.45; p &amp;lt; 0.01) and IL-8 at GFR &amp;gt; 15 ml/(min•1.73 m2) (R = 0.57; p &amp;lt; 0.01). Conclusion Systemic inflammation appears in the initial stages of CKD, being a trigger for further kidney dysfunction and the development of mineral and bone disorders. Systemic inflammation treatment in patients with CKD is staying a magnetic but still unattainable goal that needs further efforts to invest.

#1416 Extracorporeal removal of per- and polyfluoroalkyl substances (PFAS) by hemoadsorption: in vitro kinetic model

Abstract Background and Aims Per- and polyfluoroalkyl substances (PFAS) are known water pollutants leading to potential public health consequences. High blood levels of PFAS have been associated with several pathological conditions including testicular and kidney cancer. Classic extracorporeal therapies have demonstrated limited efficiency and new approaches should be explored. In this study we studied the possible role of hemoadsorption to achieve a fast, safe and effective removal of PFAS from blood in patients with high blood levels. Method We developed an in vitro model of hemoadsorption to test the potential of PFAS removal by extracorporeal treatment. We recirculated a highly polluted batch of water (4 liters) through HA380 sorbent cartridge (Jafron Medical, Zuhai, China) for 120 minutes at a flow of 150 mL/min in a closed loop configuration (Fig. 1). We collected samples at different time points and analyzed 39 different PFAS compounds. Results For the PFAS compounds with concentrations significantly above normal, we observed a removal ratio close to 90% already within the first 60 minutes of circulation leading to almost complete elimination of all pollutants at 120 minutes (Fig. 2). RR is remarkably high already in the very early moments of the adsorption process. This is likely dependent on the interaction of the different solutes with the molecular structure of the sorbent. Conclusion The in vitro model of hemoadsorption suggests the possible application in vivo of this technique to reduce/normalize the concentrations of PFAS in patients exposed to water or environmental pollution.