Routine Biochemistry Research Articles

Synthesis, biological activity evaluation and mechanism of action of novel bis-isatin derivatives as potential anti-liver cancer agents

A series of bis-isatin conjugates with lysine linker were synthesized with the aim of probing their antiproliferative potential. All the newly synthesized derivatives (0–100 μM) were first screened against liver cancer cell lines(Huh1, H22, Huh7, Hepa1-6, HepG2, Huh6 and 97H) using CCK-8 assay. Results indicated that the derivative 4d exhibited the most potent activity against Huh1 (IC50 = 17.13 µM) and Huh7(IC50 = 8.265 µM). In vivo anti-tumor study showed that compound 4d effectively inhibited tumor growth in Huh1-induced xenograft mouse model; the anti-tumor effect of compound 4d (15 mg/kg) was comparable with sorafenib (20 mg/kg). H&E staining analysis and routine blood test and blood serum biochemistry examination was performed to confirm the safety of compound 4d in xenograft models. The mechanism of action of 4d on tumor growth inhibition was further investigated by RNA-Seq analysis, which indicates a positive regulation of autophagy signaling pathway, which was further confirmed with key biomarker expression of autophagy after 4d treatment. Our results suggest that the bis-isatin conjugate compound 4d is a promising tumor inhibitory agent for some liver cancer.

<b>Bilateral phacoemulsification and intraocular lens implantation in a pet rabbit with cataracts</b>

<b>Background:</b> The occurrence of cataracts in rabbits is a common reason for consultation in veterinary ophthalmology. <b>Case Description: </b> A 6-year-old female Lop rabbit (Oryctolagus cuniculus) was referred with a 3-month history of bilateral cataracts. The owner noted that the rabbit was frequently bumping into obstacles. Physical examination was unremarkable. Routine hemogram and biochemistry panel were within usual values, and Encephalitozoon cuniculi was not detected by serology. A complete ophthalmological examination revealed bilateral hypermature cataracts causing total blindness. Bilateral tonometry showed normal intraocular pressures. Electroretinography under general anesthesia revealed strong photoreceptor responses for each eye, and ocular ultrasound was normal except for bilateral hypermature cataracts. Following electroretinography, during the same anesthesia session, conventional phacoemulsification followed by acrylic intraocular lens (IOL; +58D) implantation was performed uneventfully in both eyes. Follow-up examinations were carried out for one year. Except for immediate post-operative bilateral corneal ulcers that were quickly cured with medical treatment, no other post-operative complications were observed. The one-year follow-up examination revealed centered IOLs without posterior capsular opacity, enabling the observation of normal fundus in both eyes. The owner confirmed the rabbit had normal autonomy and moved with accuracy, as observed during follow-up visits and on videos provided by the owner. <b>Conclusion: </b> To our knowledge, this is the first described case of successful bilateral phacoemulsification and bilateral intraocular lens implantation with prior electroretinographic evaluation of the retinal function in a pet rabbit with cataracts.

The role of ischemia-modified albumin, presepsin, delta neutrophil index, and inflammatory markers in diagnosing acute cholecystitis.

The purpose of this study is to determine the significance of markers such as C-reactive protein, procalcitonin, complete blood count parameters, delta neutrophil index, ischemia-modified albumin, presepsin, and oxidative stress indicators, which are associated with inflammation, oxidative stress, and ischemia in the pathology and diagnosis of acute cholecystitis in adults. Patients diagnosed with acute cholecystitis in the emergency department and healthy individuals in the control group were included in the study. Routine blood count and biochemistry analyses were performed on the participants. Blood serum was used to measure ischemia-modified albumin, presepsin, and oxidative stress indicators. White blood cell count, neutrophil-to-lymphocyte ratio, platelet-to-lymphocyte ratio, delta neutrophil index, C-reactive protein, procalcitonin, ischemia-modified albumin, ischemia-modified albumin to albumin ratio, presepsin, and oxidative stress indicators were significantly higher in patients with cholecystitis compared to the control group. Measurements of white blood cell count, neutrophil-to-lymphocyte ratio, platelet-to-lymphocyte ratio, and delta neutrophil index can be included as part of the complete blood count. The complete blood count parameters are readily available and do not incur additional costs to the healthcare system. The authors believe that the neutrophil-to-lymphocyte ratio, delta neutrophil index, ischemia-modified albumin, ischemia-modified albumin to albumin ratio, and presepsin values can be used as new markers in the diagnosis of acute cholecystitis due to their high sensitivity, specificity, and low negative likelihood ratio.

Evaluation of the analytical performance of 80 parameters analyzed in routine biochemistry laboratory by process sigma methodology

Evaluation of the analytical performance of 80 parameters analyzed in routine biochemistry laboratory by process sigma methodology

Neurofilament Light Chain levels during admission after out-of-hospital cardiac arrest is associated with cognitive performance at 3-months follow-up

Abstract Background Patients who are resuscitated from out-of-hospital cardiac arrest (OHCA) and hospitalized in coma are at high risk of mortality or brain injury with consequent cognitive impairment, and new therapy are wanted. Biomarkers of brain injury, including Neuron-specific enolase (NSE) and Neurofilament light chain (NfL), predict mortality from cerebral injury. However, prognostication of cognitive dysfunction after discharge in OHCA survivors is challenging. Montreal Cognitive Assessment (MoCA) is a recommended standardized cognitive function testing which is used in the months following OHCA. Nevertheless, prognostic methods assessed in the initial phase of admittance which identify patients at risk of developing cognitive impairment are needed. Purpose To investigate the effect of tocilizumab administered at admission after OHCA on cognitive performance assessed by MoCA at 3-month follow-up, and to investigate the association to time-to-ROSC, and NSE as well as NfL levels during index admission with MoCA at follow-up. Methods The IMICA trial evaluated tocilizumab for reducing systemic inflammation after OHCA and randomized 80 comatose adult patients at admission to an infusion of either tocilizumab (8mg/kg, maximum 800mg) or placebo. NSE at 48 hours was measured as part of routine biochemistry based on serum samples, and NFL at 48 hours was measured in serum samples from the biobank. Time-to-ROSC was reported from pre-hospital data and cognitive performance at 3 months follow-up were assessed by the MoCA test. The association between MoCA and time-to-ROSC, NSE and NfL was assessed by Spearman’s correlation analyses. Results At follow-up, 35 patients had a MoCA test performed, while 28 patients were deceased, and 17 were alive without having an available MoCA score. There was no difference in MoCA scores at follow-up in the tocilizumab group compared to placebo group (p=0.72), and therefore results for both groups were pooled for analyses. NfL was found to be inversely correlated with MoCA score, r = -0.49, p = 0.003 (Figure), while there were no correlations with MoCA for time-to-ROSC or NSE (p = 0.46 and p = 0.34). Conclusions NfL measured at 48 hours after admission was inversely correlated with cognitive performance at 3 months for survivors of OHCA.

Abstract 12847: The Heart Bus: A Mobile Patient-Centered Approach to Improve Early Diagnosis of Heart Failure in the Community

Introduction: Heart failure with preserved ejection fraction (HFpEF) is associated with poor quality of life and considerable morbidity and mortality risk. Early diagnosis is expected to improve clinical outcomes, but requires imaging and laboratory analyses with frequent hospital visits. We developed a mobile care unit, the Heart Bus, to improve early diagnosis of HF. Methods: The Heart Bus was a 70 m 2 trailer with examination rooms, cardiac ultrasound, exercise testing equipment, a clinical laboratory, and 4 staff. During 2020-2022, the Heart Bus team reached out to the population in local newspapers, via HF patient digital platforms, and visited locations in the West Coast of Sweden. Eligible participants had suspected signs or symptoms of HF. Participants underwent echocardiography and tests for cardiopulmonary exercise, vascular function, adenosine-stress coronary flow reserve, and routine biochemistry and haematology. Molecular and physiological deep phenotyping were also performed in the Heart Bus to facilitate future patient stratification and precision medicine (data to be presented Nov 2023). Results: Of 218 participants screened over 18 months (12/month), 200 were included. Most participants actively reached out to take part. Twenty three participants (11.5%) had HFrEF, 54 (27.0%) had HFmrEF, and 62 (31.0%) had HFpEF; 70 (35.0%) had a previous HF diagnosis. Of the remaining patients, 60 (30.0%) had intermediate risk of HFpEF. The phenotypes of the EF-stratified HF subgroups were similar. However, participants in the HFpEF cohort had more women ( p < 0.001), lower lung volume ( p < 0.01), higher pulse wave velocity ( p < 0.01) and poorer kidney function ( p < 0.05) than those with HFrEF. Conclusions: The Heart Bus mobile clinic identified large numbers of patients with HF and may represent a cost-effective way to early HF diagnosis also for underserved communities, those located remotely, or have other thresholds for admittance to HF clinics.

Discovery of Taurocholic Acid Sodium Hydrate as a Novel Repurposing Drug for Intervertebral Disc Degeneration by Targeting MAPK3.

Nowadays, more than 90% of people over 50 years suffer from intervertebral disc degeneration (IDD), but there are exist no ideal drugs. The aim of this study is to identify a new drug for IDD. An approved small molecular drug library including 2040 small molecular compounds was used here. We found that taurocholic acid sodium hydrate (NAT) could induce chondrogenesis and osteogenesis in mesenchymal stem cells (MSCs). Then, an in vivo mouse model of IDD was established and the coccygeal discs transcriptome analysis and surface plasmon resonance analysis (SPR) integrated with liquid chromatography-tandem mass spectrometry assay (LC-MS) were performed in this study to study the therapy effect and target proteins of NAT for IDD. Micro-CT was used to evaluate the cancellous bone. The expression of osteogenic (OCN, RNX2), chondrogenic (COL2A1, SOX9), and the target related (ERK1/2, p-ERK1/2) proteins were detected. The alkaline phosphatase staining was performed to estimate osteogenic differentiation. Blood routine and blood biochemistry indexes were analyzed for the safety of NAT. The results showed that NAT could induce chondrogenesis and osteogenesis in MSCs. Further experiments confirmed NAT could ameliorate the secondary osteoporosis and delay the development of IDD in mice. Transcriptome analysis identified 128 common genes and eight Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways for NAT. SPR-LC-MS assay detected 57 target proteins for NAT, including MAPK3 (mitogen-activated protein kinase 3), also known as ERK1 (extracellular regulated protein kinase 1). Further verification experiment confirmed that NAT significantly reduced the expression of ERK1/2 phosphorylation. NAT would induce chondrogenesis and osteogenesis of MSCs, ameliorate the secondary osteoporosis and delay the progression of IDD in mice by targeting MAPK3.Furthermore, MAPK3, especially the phosphorylation of MAPK3, would be a potential therapeutic target for IDD treatment.

Evaluation of Arterial Stiffness in Depression Patients.

It has been known that there is a significant correlation between depression and cardiovascular diseases. However, the reasons behind this correlation that could affect mortality and morbidity were not fully identified. The present study aimed to analyze arterial stiffness diagnosed with ultrasonography, which could be associated with cardiovascular disease risks in depression patients, and to compare the findings with those of healthy controls. The study was conducted with 35 depression patients and 35 healthy individuals. Routine complete blood and biochemistry tests were requested for all patients, and their weight and height, waist circumference, and diastolic and systolic arterial blood pressure were measured. Femoral and carotid artery intima-media thickness and other arterial stiffness parameters were determined with Doppler ultrasonography. It was determined that the systolic pressure (P = .028) was higher in the patient group (P = .028). Also, the carotid elastic modulus (P = .048) was significantly higher in the patient group. A negative and significant correlation was determined between femoral compliance and chlorpromazine equivalent dose (P = .021, r = -0.389). It was determined that the systolic blood pressure and carotid elastic modulus arterial stiffness parameters were significantly higher in depression patients. Measurable arterial stiffness parameters should be investigated in depression patients as cardiovascular risk markers. Furthermore, the determination of the effects of psychotropic drugs employed in arterial stiffness treatment could play an important role in the determination of cardiovascular disease risk in these patients.

Systemic biomarkers of microvascular alterations in type 1 diabetes associated neuropathy and nephropathy - A prospective long-term follow-up study

IntroductionThis study aimed to investigate circulating biomarkers associated with the risk of developing diabetic peripheral neuropathy (DPN) and nephropathy in type 1 diabetes (T1D). Materials and methodsPatients with childhood-onset T1D (n = 49, age 38.3 ± 3.8 yrs.) followed prospectively were evaluated after 30 years of diabetes duration. DPN was defined as an abnormality in nerve conduction tests. Matrix metalloproteinase-9 (MMP-9) and its tissue inhibitor TIMP-1, neutrophil gelatinase-associated lipocalin-2 (NGAL), soluble P-selectin (sP-selectin), estimated GFR (eGFR), micro/macroalbuminuria and routine biochemistry were assessed. For comparison, control subjects were included (n = 30, age 37.9 ± 5.5 yrs.). ResultsIn all, twenty-five patients (51 %) were diagnosed with DPN, and nine patients (18 %) had nephropathy (five microalbuminuria and four macroalbuminuria). Patients with DPN had higher levels of TIMP-1 (p = 0.036) and sP-selectin (p = 0.005) than controls. Patients with DPN also displayed higher levels of TIMP-1 compared to patients without DPN (p = 0.035). Patients with macroalbuminuria had kidney disease stage 3 with lower eGFR, higher levels of TIMP-1 (p = 0.038), and NGAL (p = 0.002). In all patients, we found only weak negative correlations between eGFR and TIMP-1 (rho = −0.304, p = 0.040) and NGAL (rho = −0.277, p = 0.062, ns), respectively. MMP-9 was higher in patients with microalbuminuria (p = 0.021) compared with normoalbuminuric patients. ConclusionsOur findings indicate that TIMP-1 and MMP-9, as well as sP-selectin and NGAL, are involved in microvascular complications in T1D. Monitoring and targeting these biomarkers may be a potential strategy for treating diabetic nephropathy and neuropathy.

Multi-Omic Candidate Screening for Markers of Severe Clinical Courses of COVID-19.

Severe coronavirus disease 2019 (COVID-19) disease courses are characterized by immuno-inflammatory, thrombotic, and parenchymal alterations. Prediction of individual COVID-19 disease courses to guide targeted prevention remains challenging. We hypothesized that a distinct serologic signature precedes surges of IL-6/D-dimers in severely affected COVID-19 patients. We performed longitudinal plasma profiling, including proteome, metabolome, and routine biochemistry, on seven seropositive, well-phenotyped patients with severe COVID-19 referred to the Intensive Care Unit at the German Heart Center. Patient characteristics were: 65 ± 8 years, 29% female, median CRP 285 ± 127 mg/dL, IL-6 367 ± 231 ng/L, D-dimers 7 ± 10 mg/L, and NT-proBNP 2616 ± 3465 ng/L. Based on time-series analyses of patient sera, a prediction model employing feature selection and dimensionality reduction through least absolute shrinkage and selection operator (LASSO) revealed a number of candidate proteins preceding hyperinflammatory immune response (denoted ΔIL-6) and COVID-19 coagulopathy (denoted ΔD-dimers) by 24-48 h. These candidates are involved in biological pathways such as oxidative stress/inflammation (e.g., IL-1alpha, IL-13, MMP9, C-C motif chemokine 23), coagulation/thrombosis/immunoadhesion (e.g., P- and E-selectin), tissue repair (e.g., hepatocyte growth factor), and growth factor response/regulatory pathways (e.g., tyrosine-protein kinase receptor UFO and low-density lipoprotein receptor (LDLR)). The latter are host- or co-receptors that promote SARS-CoV-2 entry into cells in the absence of ACE2. Our novel prediction model identified biological and regulatory candidate networks preceding hyperinflammation and coagulopathy, with the most promising group being the proteins that explain changes in D-dimers. These biomarkers need validation. If causal, our work may help predict disease courses and guide personalized treatment for COVID-19.

Experimental infection and co-infection with Chinese strains of Ehrlichia canis and Babesia vogeli in intact and splenectomized dogs: Insights on clinical, hematologic and treatment responses

Animal infection models are crucial for studying various aspects of Ehrlichia canis infections. To understand the pathogenesis of the first Chinese isolate of E. canis and simulate the natural progression of canine ehrlichiosis, we developed a model with 18 Beagle dogs that consisted of E. canis initial infection (days 0–17), treatment with doxycycline or rifampicin (days 18–32), recovery (days 33–66), E. canis reinfection (days 67–91), and Babesia vogeli superinfection (days 92–116). We measured body weight and rectal temperature every other day, drew blood every 4 days for routine hematology and biochemistry tests, and for quantification of E. canis and B. vogeli by quantitative PCRs. In this study, the first isolate of E. canis from China was used to experimentally infect dogs, and the infected dogs exhibited clinical signs of acute severe ehrlichiosis, including high fever, loss of appetite, dehydration, and body weight loss, confirming the similar pathogenicity of E. canis in China as compared to isolates from other regions. Infection with E. canis and B. vogeli led to reduced body weight and fever in dogs. Doxycycline treatment led to absence of E. canis DNA in infected dogs, while rifampicin treatment lowered the blood E. canis copy number up to 1.5 folds. E. canis-free infected dogs after doxycycline treatment were successfully re-infected with E. canis, indicating dogs with antibodies are still at risk of re-infection. Super-infection with B. vogeli resulted in higher fever, more severe anemia, and a reduced number of platelets. Splenectomized dogs showed significantly higher E. canis numbers during recovery and re-infection than intact dogs. The histological changes were observed in brain, lung, kidney, liver and spleen of the infected dogs. The findings in this study provide insights into clinical and hematologic responses, as well as effective treatment options, for dogs infected with the first Chinese isolate of E. canis, and may contribute to our understanding of the diagnosis and prevention of tick-borne diseases in dogs, including canine monocytic ehrlichiosis.

Parameters Predicting Microvascular Invasion and Poor Differentiation in Hepatocellular Carcinoma Patients with Normal Alpha-fetoprotein Level Before Liver Transplantation.

The aim of this study is to evaluate the parameters that might be associated with pathologically diagnosed microvascular invasion and poor differentiation, using complete blood count and routine clinical biochemistry test results, in hepatocellular carcinoma patients before liver transplantation. The data of patients who underwent liver transplantation for hepatocellular carcinoma at our institute, between March 2006 and November 2021, was researched retrospectively. The incidence of microvascular invasion was 28.6%, poor differentiation rate was 9.3%, hepatocellular carcinoma recurrence rate after liver transplantation was 12.1%, and median time to recurrence was 13 months, in the patients with normal alpha-fetoprotein levels. After univariate and multivariate analysis, maximum tumor diameter > 4.5 cm and the number of nodules (n > 5) were found to be independent risk factors for microvascular invasion, and number of nodules > 4 and mean platelet volume ≤8.6 fL were found to be independent risk factors for poor differentiation. Serum alpha-fetoprotein levels were still within the normal range at the recurrence time, in 53% of the patients who had recurrence after liver transplantation, but surprisingly were elevated in 47% of the patients at time of hepatocellular carcinoma recurrence. In hepatocellular carcinoma patients with normal alpha-fetoprotein levels before liver transplantation, independent risk factors of the presence of microvascular invasion were maximum tumor diameter and number of nodules, and independent risk factors of poor differentiation were mean platelet volume and number of nodules. Furthermore, serum alpha-fetoprotein levels were still normal at time of recurrence in 53% of hepatocellular carcinoma patients whose alpha-fetoprotein levels were normal before liver transplantation but were elevated in 47% of the patients at recurrence time, despite having normal levels before liver transplantation.

Plasma lipoprotein subclass variation in middle-aged and older adults: Sex-stratified distributions and associations with health status and cardiometabolic risk factors

Circulating lipids and lipoproteins mediate cardiovascular risk, however routine plasma lipid biochemistry provides limited information on pro-atherogenic remnant particles. We analysed plasma lipoprotein subclasses including very low-density and intermediate-density lipoprotein (VLDL and IDL); and assessed their associations with health and cardiometabolic risk. From 1,976 community-dwelling adults aged 45-67 years, 114/1071 women (10.6%) and 153/905 men (16.9%) were categorised as very healthy. Fasting plasma lipoprotein profiles comprising 112 parameters were measured using 1H nuclear magnetic resonance (NMR) spectroscopy, and associations with health status and cardiometabolic risk factors examined. HDL cholesterol was higher, and IDL and VLDL cholesterol and triglycerides lower, in very healthy women compared to other women, and women compared to men. IDL and VLDL cholesterol and triglyceride were lower in very healthy men compared to other men. HDL cholesterol and apolipoprotein (apo) A-I were inversely, and IDL and VLDL cholesterol, apoB-100, and apoB-100/apoA-I ratio directly associated with body mass index (BMI) in women and men. In women, LDL, IDL and VLDL cholesterol increased with age. Women with diabetes and cardiovascular disease had higher cholesterol, triglycerides, phospholipids and free cholesterol across IDL and VLDL fractions, with similar trends for men with diabetes. Lipoprotein subclasses and density fractions, and their lipid and apolipoprotein constituents, are differentially distributed by sex, health status and BMI. Very healthy women and men are distinguished by favorable lipoprotein profiles, particularly lower concentrations of VLDL and IDL, providing reference intervals for comparison with general populations and adults with cardiometabolic risk factors.

Analysis of Ischemia-Modified Albumin (IMA) and Coagulation Parameters in Patients with SARS-CoV-2 Pneumonia

Background: Coronavirus disease 2019 (COVID-19) is a systemic disease which causes an increased inclination to thrombosis by leading to coagulation system activation and endothelial dysfunction. Our objective in this study is to determine whether ischemia-modified albumin (IMA) can be used as a new marker in patients with COVID-19 for evaluating the increased coagulation risk, pneumonic infiltration, and thus, prognosis. Methods: Our study included 59 patients with COVID-19 compatible pneumonic infiltration on lung computed tomography (CT) who applied to and were hospitalized in the Internal Diseases Outpatient Clinic, then followed up and treated, as well as 29 healthy individuals with a negative COVID-19 rRT-PCR test without any additional disease. Hemogram, coagulation, routine biochemistry, and serum IMA activity parameters were studied. Results: In our study, the higher serum IMA level in COVID-19 patients with pneumonic infiltration compared to that of the healthy control group was found to be statistically significant. No significant correlation was found between the serum IMA levels and the coagulation and inflammation parameters in the 59 COVID-19 patients included. Conclusions: Serum IMA levels in COVID-19 patients with pneumonic infiltration on CT were found to be higher than in the control group. Examination of biochemical parameters, especially thrombotic parameters that affect prognosis such as IMA, can be a guide in estimating pneumonic infiltration.

Hypophosphataemia definitions, incidence and associated outcomes in paediatric intensive care: A retrospective cohort study in post-cardiac surgical patients

Hypophosphataemia has been linked to higher morbidity and mortality in intensive care but there is inconsistency in the definition of hypophosphataemia for infants and children. We aimed to determine the incidence of hypophosphataemia in a group of at-risk children in paediatric intensive care unit (PICU) and associations with patient characteristics and clinical outcomes using three different hypophosphataemia thresholds. Retrospective cohort study of 205 post-cardiac surgical patients <2 years of age admitted to Starship Child Health PICU, Auckland, New Zealand. Patient demographics and routine daily biochemistry for 14 days after PICU admission were collected. Rates of sepsis, mortality and length of mechanical ventilation were compared between groups with different serum phosphate concentrations. Out of 205 children, 6 (3%), 50 (24%) and 159 (78%) had hypophosphataemia at thresholds of <0.7, <1.0 and <1.4 mmol/L, respectively. There were no differences in gestational age at birth, sex, ethnicity or mortality in those with and without hypophosphataemia at any threshold. Children with a serum phosphate <1.4 mmol/L had more mean (SD) total hours of mechanical ventilation (85.2 (79.6) vs. 54.9 (36.2) h, P = 0.02) and those with mean serum phosphate <1.0 mmol/L had more mean hours of mechanical ventilation (119.4 (102.8) vs. 65.2 (54.8) h, P < 0.0001), episodes of sepsis (14% vs. 5%, P = 0.03) and longer length of stay (6.4 (4.8-20.7) vs. 4.9 (3.9-6.8) days, P = 0.02). Hypophosphataemia is common in this PICU cohort and serum phosphate <1.0 mmol/L is associated with increased morbidity and length of stay.

Coffee intake one hour prior to phlebotomy produces no clinically significant changes in routine biochemical test results.

Although current guidelines recommend not drinking coffee prior to phlebotomy, our hypothesis is that drinking coffee does not affect the clinical interpretation of biochemical and haematological test results. Twenty-seven volunteers were studied in basal state (T0) and 1h after (T1) drinking coffee. Routine haematological (Sysmex-XN1000 analyser) and biochemistry parameters (Vitros 4600 analyser) were studied. Results were compared using the Wilcoxon test (P < 0.05). A clinical change was considered when mean percent difference (MD%) was higher than the reference change value (RCV). Coffee intake produced statistically, but not clinically, significant: i) increases in haemoglobin (P = 0.009), mean cell haemoglobin concentration (P = 0.044), neutrophils (P = 0.001), albumin (P = 0.001), total protein (P = 0.000), cholesterol (P = 0.025), high density lipoprotein cholesterol (P = 0.007), uric acid (P = 0.011), calcium (P = 0.001), potassium (P = 0.010), aspartate aminotransferase (P = 0.001), amylase (P = 0.026), and lactate dehydrogenase (P = 0.001), and ii) decreases in mean cell volume (P = 0.002), red cell distribution width (P = 0.001), eosinophils (P = 0.002), and lymphocytes (P = 0.001), creatinine (P = 0.001), total bilirubin (P = 0.012), phosphorus (P = 0.001), magnesium (P = 0.007), and chloride (P = 0.001). Drinking a cup of coffee 1 hour prior to phlebotomy produces no clinically significant changes in routine biochemical and haematological test results.

#3444 HOW WELL INFORMED ARE PATIENTS ABOUT DIALYSIS INITIATION?

Abstract Background and Aims Studies indicate that 30% of patients on dialysis feel inadequately informed about various dialysis options and also lack involvement in the decision-making process surrounding dialysis initiation. The aim of this study was to 1) describe the provision of information to older patients with chronic kidney disease before KRT and 2) to study the association between the perceived amount of information received before dialysis and patient satisfaction with treatment post dialysis initiation. Method The EQUAL study is a multicenter, prospective cohort study in six European countries (Germany, Italy, Netherlands, Poland, Sweden, and United Kingdom). Inclusion criteria are older adults &amp;gt;65 years with an incident eGFR &amp;lt;20 ml/min/1.73 m2. Patients were followed up every 3-6 months and received routine medical care as provided by the nephrology clinic in each country. Standardized data were collected at each visit, including demographics, lifestyle, comorbidities, uremic signs and symptoms, quality of life, treatment satisfaction according to renal treatment satisfaction questionnaire (RTSQ-11), nutritional status, medication and routine blood and urine biochemistry. We described patient characteristics and perceived information provision at different time points (incident eGFR&amp;lt;20, eGFR&amp;lt;15, eGFR&amp;lt;10 ml/min/1.73 m2). Subsequentially, we compared patient characteristics between those who reported receiving more versus less information at the last eGFR before start of KRT (or end-of follow-up in patients with eGFR &amp;lt;10 ml/min/1.73 m2). We used multivariable logistic regression models to investigate if perceived information provision before start of dialysis was associated with a treatment satisfaction. Results We included 1372 patients at baseline with a median age of 76 (IQR 70–81), 34% (n = 462) women. Over follow-up, 997 of these patients progressed to an eGFR&amp;lt;15, and 573 to an eGFR&amp;lt;10 ml/min/1.73 m2. The proportion of patients who reported having received enough information about the decision on when to start dialysis increased with decreasing eGFR (53.2% at baseline to 61.2% (eGFR&amp;lt;15) and 70.9% (eGFR&amp;lt;10)). Among the 638 patients progressing to KRT or eGFR &amp;lt;10 ml/min/1.73 m2, the mean eGFR was 8.9 ml/min/1.73 m2 (SD 3.4) at their last pre-dialysis visit. Patients who reported that they had received sufficient information regarding dialysis timing were in general younger, where median age was 74 (IQR 69.4-79.3) years for those receiving information and 76 (IQR 70.7-81.4) years for those perceiving not having enough information, but there were otherwise no major differences in patient characteristics. Among these patients, 32% had still not decided on which dialysis modality to start with. There was not any statistically significant difference in RSTQ-11 score associated with type of dialysis modality (hemodialysis versus peritoneal dialysis) post dialysis initiation. Among 302 patients who started dialysis, those reporting to have received more information (highest quartile) before start of dialysis also reported being more satisfied with their treatment post dialysis (OR: 1.22; 95% CI 1.05-1.37). Conclusion Although most patients report having received pre-dialysis information, many patients with low kidney function still perceive not having received enough information to make an informed decision. Patients who are well informed are more satisfied with their treatment.

Plasma metabolomics identifies metabolic alterations associated with the growth and development of cat.

The purpose of our study was to study the composition and content of the feline plasma metabolome revealing the critical metabolites and metabolic pathways associated with age during growth and development. Blood samples were collected from juvenile and adult groups for blood routine tests and serum biochemistry tests. Non-targeted metabolomics analyses of plasma were also performed to investigate changes in metabolites and metabolic pathways. In this study, we found that the red blood cell counts, liver function indexes (albumin and gamma-glutamyl transpeptidase), and the concentration of triglyceride and glucose changed significant with growth and development. The metabolomics results revealed that 1427 metabolites were identified in the plasma of young and adult cats. Most of these metabolites belong to major classes of lipids and lipid-like molecules. The most obvious age-related metabolites include reduced levels of chenodeoxycholate, taurocholate, cholate, and taurochenodeoxycholate but increased levels of L-cysteine and taurocyamine in the adult cat's serum. These metabolites are mainly involved in the primary bile acid biosynthesis pathway, the bile secretion pathway, and the taurine and hypotaurine metabolism pathway. This study revealed many age-related metabolite alterations in the feline plasma. These age-varying metabolites, especially in the bile acid biosynthesis and secretion metabolism pathways, indicate that the regulation of these pathways is involved in the growth and development of cats. This study promotes our understanding of the mechanism of feline growth and provides new insights into nutrition and medicine for cats of different ages.

The Importance of Extended High Frequencies in Hearing Evaluation of Pediatric Patients with Type 1 Diabetes.

Type 1 diabetes (T1D), one of the most common childhood diseases worldwide, can cause hearing loss through its systemic effects. Diabetes-induced hearing loss is considered a progressive sensorineural hearing loss with a gradual onset typically occurring at high frequencies. We investigated extended high-frequency (EHF) hearing sensitivity in children with T1D who did not complain of hearing loss as an early marker for hearing loss at the standard/conventional frequency range of hearing. We evaluated 42 children (21 with T1D and 21 healthy controls) in a case-control design. Conventional and EHF (14,000, 16,000, and 18,000 Hz) audiometry were performed. The diabetes group underwent routine blood biochemistry and glycated hemoglobin (HbA1c) measurements. The data were analyzed by the Student t-test, Mann-Whitney U test, chi-square test, and logistic regression analysis. The mean hearing thresholds were significantly higher (p < 0.05) in the diabetes group than in controls at 500, 2,000, 4,000, and 8,000 Hz (all < 15 decibel hearing level [dB HL]). The number of ears with thresholds > 15 dB HL at 14,000-18,000 Hz but ≤ 15 dB HL at 500-4,000 Hz was significantly higher in the diabetes group than in the control group (p = 0.049). Children with diabetes showed normal hearing thresholds within the conventional audiometric frequency range; however, they presented higher hearing thresholds during EHF audiometry when they were compared with controls. Audiometry in these children should be performed using frequencies above 8,000 Hz combined with the conventional frequency range. EHF audiometry could be an effective method for identifying subclinical hearing loss in children with diabetes. Thus, diabetic children with an EHF mean hearing threshold above 15 dB HL should be monitored more closely in terms of blood glucose regulation to prevent diabetes-related hearing loss at the conventional frequency range.

Retinochoroidal Manifestation of Toxoplasmosis in a Young Patient

We report the case of a 15-year-old young girl, with retinochoroidal manifestation of toxoplasmosis, without vitritis. Serology for Toxoplasma gondii was positive (IgG positive /IgM negative), but PCR for Toxoplasma gondii in peripheral blood samples was negative. Blood tests including complete hemogram (total leukocyte, red blood cell and platelet count) except for hemoglobin, coagulogram (APTT, INR, and fibrin degradation product level) and routine blood biochemistry (serum urea, creatinine, and random blood sugar) were within normal limits. The serum levels of triglycerides, low density lipoproteins, high-density lipoproteins and calcium were within normal limits. Direct ophthalmoscopy of the right eye revealed a well-demarcated circular atrophic pigmented scar of about four disc diameters involving the macula. We did not initiate any therapy, because the process appeared to be active anymore. In our case demonstrate of retinochoroidal manifestation of toxoplasmosis by routine fundus examination in a young patient with uncharacteristic complaints.