Assessing the status of European laboratories in evaluating biomarkers for chronic kidney diseases (CKD) and recommendations for improvement: insights from the 2022 EFLM Task Group on CKD survey.

OBJECTIVES: Chronic kidney disease (CKD) is a global health issue, ranking as the third leading cause of death worldwide. CKD diagnosis and management depend on clinical laboratory tests, necessitating consistency for precise patient care. Global harmonization of CKD testing through clinical practice guidelines (CPGs) is recommended. Prior to CPG development, assessing the current CKD testing landscape is crucial. In 2022, the European Federation of Laboratory Medicine (EFLM) conducted an online survey among European laboratories associated with EFLM, evaluating CKD testing practices, including new glomerular filtration rate (GFR) estimation methods. This report summarizes the 2022 survey findings and offers recommendations for improving CKD test standardization. METHODS: An online survey was conducted in November 2022 using a questionnaire hosted on LimeSurvey sent to European laboratories affiliated with the EFLM. The survey results were recorded in Excel files and analysed. RESULTS: The results highlight significant discrepancies among countries in unit expression, methods, cystatin C use, and GFR calculation equations. Additionally, limited attention to pediatric renal biology specifics, varied proteinuria and albuminuria result expressions, and limited awareness of GFR measurement methods through iohexol clearance are noted. CONCLUSIONS: In an effort to enhance the standardization of crucial biomarkers utilized in nephrology for evaluating renal function and diagnosing kidney injuries, the EFLM Task Group on CKD suggests nine practical recommendations tailored for European laboratories. The group is confident that implementing these measures will minimize result expression discrepancies, ultimately leading to enhanced patient care.

Cystic fibrosis related bone disease in children: Can it be predicted?

BACKGROUND&AIMS: Cystic fibrosis (CF) -related bone disease (CFBD) is an important complication of CF, and low BMD in childhood is a precursor of CFBD. Here, we aimed to investigate bone turnover biomarkers, including osteocalcin (OC), receptor activator of nuclear factor kappa B ligand (RANKL) and osteoprotegerin (OPG) in relation to low BMD in children with CF (cwCF). We also evaluated factors which could affect bone turnover with particular emphasis on fat-free mass (FFM), forced expiratory volume in 1 s (FEV1), hand grip strength (HGS), and functional capacity and physical activity. METHODS: Sixteen cwCF aged 8-18 years with moderate low BMD (group1) and 64 cwCF with normal BMD (group2) were enrolled. Serum RANKL, OC, and OPG were determined by immunoenzymatic assays. Multiple parameters including pancreatic status, lung functions, body mass index (BMI), FFM measured by bioelectric impedance analysis (BIA), 6-minute walk test, vitamin D, nutritional intake, HGS, functional capacity and physical activity, serum and urine biomarkers were compared between the two groups. RESULTS: We found similar serum levels of RANKL (p = 0.501), OC (p = 0.445), OPG (p = 0.380), and RANKL/OPG ratio (p = 0.449) between group1 and group2 in cwCF. BMI z-score (p < 0.001), FFMI z-score (p < 0.001), FEV1 z-score (p = 0.007), and right-HGS (%pred) (p = 0.009) significantly differed between the two groups. Multivariate linear regression revealed that the only factors that predicted BMD were FFMI z-score and HGS %pred. CONCLUSION: Serum OC, OPG, RANKL and RANKL/OPG ratio did not predict BMD in cwCF. FFMI z-score and HGS %pred measured by non-invasive and practical methods were the best predictors of BMD.

Interleukin-22/Interleukin-22 binding protein axis and oral contraceptive use in polycystic ovary syndrome.

PURPOSE: Polycystic ovary syndrome (PCOS) is associated with alterations in gut microbiota. The cytokine interleukin-22 (IL-22) is produced by immune cells and closely linked to gut immunity, which is tightly controlled by its binding protein (IL-22BP). In this study, we aimed to assess whether IL-22/IL-22BP axis is altered in PCOS at baseline and in response to short-term oral contraceptive (OC) therapy. METHODS: We have evaluated circulating concentrations of IL-22 and IL-22BP in serum samples of 63 PCOS patients and 39 age- and BMI-matched healthy controls. Blood samples were taken in the early follicular phase of a cycle and stored at -80 °C. Serum IL-22 and IL-22BP levels were measured by ELISA at baseline in both women with PCOS and controls, and after 3 months of OC use in PCOS group. IL-22/IL-22BP ratio was calculated in order to have a better reflection of IL-22 biological activity. RESULTS: At baseline, serum IL-22, IL-22BP concentrations and IL22/IL-22BP ratio were similar between women with PCOS and healthy controls. Three months of OC use along with general lifestyle advice resulted in a significant increase in IL-22/IL-22BP ratio in the PCOS group (62.4 [IQR:14.7-172.7] at baseline vs 73.8 [IQR:15.1-264.3] after OC use respectively p = 0.011). CONCLUSIONS: Results of this study show that women with PCOS have similar circulating concentrations of IL-22 and IL-22BP with healthy women and that short term oral contraception is associated with an increase in IL-22/IL-22BP ratio suggesting higher biological activity of the IL-22 system with OC use in PCOS.

Effect of anti-epileptic drugs on first trimester screening test results.

OBJECTIVE: To evaluate first trimester screening test parameters in epileptic patients using anti-epileptic drugs. MATERIALS AND METHODS: We retrospectively evaluated first trimester screening test results of 23 epileptic pregnant women using anti-epileptic drugs with a control group consisting of 92 healthy pregnancies. The anti-epileptic drugs used in this study were carbamazepine, levatiracetam, valproic acid and lamotrigine. Single drug or multi-drug regimens were used according to the clinical conditions. Patients with any known chronic or acute disease and drug usage were excluded from the study. Comparisons were performed via Mann-Whitney U test. RESULTS: First trimester screening test biochemical markers were compared and maternal serum PAPP-A MoM values were found to be similar in study and control groups while ß-hCG MoM values were significantly higher in pregnancies using epileptic drugs (p: 0,737 and p < 0.001, respectively). CONCLUSION: Biochemical first trimester screening test results may be affected by anti-epileptic drug usage, which may lead to misinterpretation of the risk level. Thus, validation of MoM values should be necessary in order to obtain optimal results.

Comparison of term and preterm labor procalcitonin and leukocyte cell volume, conductivity and light scatter (VCS) parameters in order to demonstrate the impact of inflammation on the triggering mechanisms of preterm uterin contractions.

AIM: Comparison of mean channels of cell volume, conductivity and light scatter (VCS) parameters of neutrophil, monocyte and lymphocyte, procalcitonin (PCT) and white blood cell count (WBC) during term and preterm labor to evaluate the impact of inflammation on the triggering mechanisms of uterine contractions. METHODS: This study is consisted of 16 preterm and 60 term pregnancies at the beginning of the first stages of the labor. Leukocyte VCS parameters, PCT plasma levels and WBC count were evaluated. RESULTS: We could not demonstrate statistically significant difference in between leukocyte VCS parameters in preterm and term deliveries (P ˃ 0.050 for all). WBC counts were 10.6 and 11.8 × 103 /µL in the preterm and term groups respectively (P = 0.270). PCT levels were 0.04 and 0.03 ng/mL for preterm and term pregnancies (P = 0.062). CONCLUSION: Inflammation related markers such as leukocyte VCS parameters, PCT values and WBC count does not differentiate at the first stage of labor in preterm and term deliveries. These variables do not seem to have a prominent role at the biological events behind preterm contractions.

Is muscle mechanical function altered in polycystic ovary syndrome?

PURPOSE: Polycystic ovary syndrome (PCOS) is the most common endocrine disorder of women of reproductive age. The aim of the current study was to assess muscle mechanical function in PCOS and its relationship with hormonal and metabolic features of the syndrome. METHODS: The study included 44 women with PCOS, all having clinical or biochemical hyperandrogenism, chronic oligo-anovulation and PCOM, and 32 age- and BMI-matched healthy women. Anthropometric, hormonal and biochemical measurements were performed. Muscle mechanical function including lower limb explosive strength and average power (AvP) was measured using isokinetic dynamometry, a valid and reliable instrument for measuring muscle strength. RESULTS: The mean age and BMI of the women with PCOS and controls were 21.8 ± 3.2 versus 22.8 ± 3 years and 26.1 ± 5.4 versus 25.5 ± 5.7 kg/m2, respectively (p = NS for both). PCOS patients had higher androgen levels, whereas total and regional fat and lean body mass and insulin resistance parameters were similar between the groups. The peak muscle force output defined as the peak torque of knee extensor and flexor muscles was higher in normal weight women compared to overweight and obese (p < 0.05 for both) but did not differ in patients and controls. AvP determined by the time-averaged integrated area under the curve at 60°/s angular velocity was higher in the PCOS group for extension and flexion (50.3 ± 21.2 vs 42.1 ± 11.6 and 35.3 ± 27 vs 22.2 ± 11.1, respectively, p < 0.05 for both). These measurements were correlated with bioavailable testosterone (r = 0.29, p = 0.012, r = 0.36, p = 0.001, respectively). CONCLUSION: Muscle mechanical function is altered in PCOS. Women with PCOS have increased average lower limb power that is associated with hyperandrogenism.

Human papillomavirus infection and autoimmune disorders: a tertiary center experience.

This study aimed to investigate the relationship between HPV and autoimmune disorders. We retrospectively evaluated 62 women who had HPV-DNA positivity in terms of autoimmune disorders (autoimmune antibody positivity, chronic inflammatory diseases and autoimmune diseases). The patients were divided into two groups according to autoimmune disorder positivity (autoimmune positive (n = 30), autoimmune negative (n = 32)) and compared with each other in terms of single and multiple HPV-DNA types, high and low-risk HPV-DNA types, and Pap smear findings. We determined that 48.4% of the HPV-DNA positive patients had autoimmune disorders. We found that 15 of 62 (24.2%) women had more than one type of HPV and HPV type 16 was the dominant type in this study (58.2%). A total of 27.4% of HPV-DNA positive patients had abnormal cytological findings. There was no statistically significant difference between autoimmune groups in terms of the presence of high-risk HPV types, multiple HPV types and abnormal cytological findings (P = 0.531, P = 0.558 and P = 0.234, respectively). The prevalence of autoimmune disorders was high among HPV-DNA positive women. On the other hand, the rate of high-risk HPV type positivity, multiple HPV infections and cytopathological findings were similar between the autoimmune positive and negative groups.

Emerging biomarker for predicting acute kidney injury after cardiac surgery: cystatin C

Background/aim: Cardiopulmonary bypass (CPB)-associated acute kidney injury (AKI) is a common situation and rapid diagnosis and risk classification are important in the prevention and management of AKI. Changes in serum creatinine (SCr) levels in the current consensus criteria do not allow clinicians to diagnose CPB-associated AKI until 48 h after surgery. Materials and methods: We conducted a prospective single center study involving 57 patients who underwent cardiac surgery with CBP to compare serum beta-trace protein (BTP), neutrophil gelatinase-associated lipocalin-2 (NGAL), and cystatin C (CysC) levels with SCr for early diagnosis of CPB associated AKI. We defined AKI according to KDIGO criteria. Results: AKI was diagnosed in 24 (42.1%) patients. Mean duration of postoperative intensive care unit stay was 4.79 (± 6.12) days for the AKI+ group and 2.15 (± 0.56) days for the AKI­ group. The average length of hospital stay was 8.13 (± 5.75) days for the AKI+ group and 7.21 (± 3.68) days for the AKI­ group, which was significantly longer in the AKI+ group (P < 0.001, P = 0.011). Unlike other biomarkers, a significant increase in postoperative CysC levels was always found in patients with AKI during follow-up when compared to patients in whom AKI did not develop (P < 0.001). Conclusion: Increase in serum CysC levels showed a significant positive correlation with increase in SCr levels. We have not seen this correlation between other biomarkers and SCr. According to our study, serum CysC was a reliable biomarker that may aid in the early detection and follow-up of AKI after cardiac surgery.

Factors associated with pulmonary hypertension and long-term survival in bronchiectasis subjects.

BACKGROUND: The development of pulmonary hypertension (PH) and its effect on long-term survival in bronchiectasis subjects has not been explored. The present study aims to analyze the factors associated with PH and its effect on long-term survival in bronchiectasis subjects. METHODS: We prospectively evaluated 23 bronchiectasis subjects without PH and 16 with PH, as well as 20 healthy volunteers. RESULTS: Bronchiectasis subjects with PH were more hypoxemic and had a greater number of involved lobes in high resolution computed tomography (HRCT) than did the bronchiectasis subjects without PH (P < 0.001 and P < 0.001, respectively). At three years, the survival rate was 95.7% for bronchiectasis subjects without PH and 56.3% for bronchiectasis with PH, and at 5 years, these rates were 95.7% and 62.5%, respectively (P = 0.002). Multivariate Cox regression analysis revealed that only the Medical Research Council (MRC) dyspnea score was independently related to poor survival in all bronchiectasis subjects (hazard ratio: 6.98; 95% CI: 2.41-20.23; P < 0.00001). CONCLUSIONS: Subjects with PH are more hypoxemic and have a greater number of involvements in the lobes of the lungs. Bronchiectasis subjects with PH have worse survival than do bronchiectasis subjects without PH. MRC dyspnea score is an independent predictor of long-term survival.

Efficacy of new leukocyte parameters versus serum C-reactive protein, procalcitonin, and interleukin-6 in the diagnosis of neonatal sepsis.

BACKGROUND: The aim of this study was to investigate and compare the efficacy of the new leukocyte parameters mean neutrophil and monocyte volume (MNV, MMV), conductivity (MNC, MMC), scattering (MNS, MMS) and volume distribution width (NDW, MDW) with serum C-reactive protein (CRP), procalcitonin (PC) and interleukin (IL)-6 in the diagnosis of neonatal sepsis. METHODS: A total of 227 newborns (132 boys, 95 girls) were analyzed. There were 116 infants in the sepsis group (proven sepsis, n = 40; clinical sepsis, n = 76) and 111 in the control group. Venous blood samples were collected from infants at the time of diagnosis and complete blood count, peripheral blood smear, blood cultures, CRP, PC, IL-6 and MNV, MMV, MNC, MMC, MNS, MMS, NDW, and MDW were analyzed. RESULTS: MNV, NDW, MMV and, MDW were higher in infants with sepsis than in controls (P < 0.05 for all). MNS was lower in the patients with sepsis (P = 0.002). There was no significant difference between the sepsis and control groups in terms of MNC, MMC and MMS. CONCLUSION: Although the predictive value of leukocyte parameters including neutrophil and monocyte volume, conductivity, scattering and volume distribution width in the diagnosis of neonatal sepsis was lower than that of CRP, PC and IL-6, some of these new parameters may be useful in the differential diagnosis of newborn sepsis, along with the other screening tools. In particular, MNV seems to be the most useful parameter with the highest specificity; also, the importance of PC in the diagnosis of early onset sepsis was confirmed.

Extremely high parathyroid hormone concentrations associated with pityriasis rubra pilaris and monoclonal gammopathy of unknown significance: a clinical dilemma.

We present a case with extremely high parathyroid hormone (PTH) concentrations in the order of hundred thousands accompanied by dermatological and hematological diseases. After several diagnostic interventions, no malignancy could be demonstrated except monoclonal gammopathy of unknown significance. The dermatological findings were taken to be manifestations of the hematological disease. Since the first serum intact PTH concentration of the patient was found to be higher than 2500 pg/ml, dilution study was performed and found to be 215,977 pg/ml. The high concentration of serum PTH was taken to be falsely high due to assay interference. This concentration was checked from three different paths; a test for linear dilution was performed, the test was repeated with another method and the sample was treated to remove or inhibit interfering substances. The results were compatible with endogenous antibody interference, presumed to be a result of monoclonal gammopathy. The extremely high PTH concentrations were not only due to assay interference, but also secondary hyperparathyroidism, which was evident by the decrease in PTH concentrations with calcium and vitamin D treatments.

Determination of circulating tumor cells for detection of colorectal cancer progression or recurrence.

The role of circulating tumor cells (CTCs) in the management of colorectal cancer has not yet established. In present study, we aimed to investigate the relationship between clinicopathological characteristics and CTC levels in patients with colorectal carcinoma. The isolation of CTCs in blood was performed with the semi-automated CellSearch System (Veridex TM). Ninety-three patients were analyzed. Median age of patients at the time of diagnosis was 56 (24-78). Total number of blood samples collected for CTCs were 130. CTCs were detected in 46 patients (> or =1 per 7.5 mL), but only thirty (32%) patients were interpreted as positive (> or = 3/7.5 mL). In the 27 patients in whom 2 blood samples were taken, levels of CTCs were found to be decreased in 8 patients, increased in 14 patients and did not change in 5 patients after therapy. Although only 2 (25%) of 8 patients in whom CTCs decreased after therapy had disease progression, 13 (93%) of 14 patients with increased CTCs had disease progression (p = 0.001). Similarly, 4 (80%) of 5 patients with stable CTCs had also disease progression. Our findings indicate that decreased CTCs after therapy is related with remission while increased CTC level is associated with disease recurrence or progression.

Construction of recB-recD genetic fusion and functional analysis of RecBDC fusion enzyme in Escherichia coli.

BACKGROUND: recD, located between recB and argA, encodes the smallest polypeptide (60 kDa) of the heterotrimeric enzyme RecBCD in Escherichia coli. RecD is a 5'-3' helicase and is required for the nuclease activity of RecBCD and for tight binding to dsDNA ends. Here, we have tested the hypothesis that RecD regulates the structure and activities of RecBCD, including RecA loading. RESULTS: To characterize its regulatory functions, recD was genetically fused to recB through deletion and substitution mutations. The recB-recD fusion led to a decreased amount of the heterotrimer. Both fusion mutants proved to be recombination proficient, viable and resistant to DNA damaging agents, and to have DNA unwinding, ATP-dependent dsDNA exonuclease and Chi genetic activities. CONCLUSION: Our findings suggest that the recB-recD fusion may form a RecBD fusion protein and therefore affect RecD assembly, but this does not change the three-dimensional structure of the heterotrimer.

Serum troponin T levels and echocardiographic evaluation in children treated with doxorubicin.

OBJECTIVE: We investigated the usefulness of serum cardiac Troponin T (cTnT) to detect doxorubicin related cardiotoxicity as a non-invasive and reliable method. PATIENTS AND METHODS: Twenty-four patients who received doxorubicin for their solid tumors at cumulative doses of 400 mg/m(2) or higher, between June 1982 and August 2000, were included in this study. None of them had clinical signs or symptoms of cardiotoxicity. The age range was 3-31 years (median 14), and male to female ratio was 14/10. The systolic and diastolic cardiac functions were evaluated by two-dimensional, M-mode, and Doppler echocardiography. Serum cTnT levels were measured by a third generation immunoassay method and the lowest detectable level was 0.010 ng/ml. RESULTS: The cumulative doxorubicin doses were at the range of 400 and 840 mg/m(2) (median 480). The time past from the last doxorubicin dose was 1-168 months (median 12). All of the patients had normal chest X-rays, electrocardiograms, and nine patients (37.5%) had abnormal systolic or diastolic cardiac function parameters. The median cumulative doxorubicin doses of the patients with normal and abnormal echocardiographic parameters were 480 and 440 mg/m(2), respectively. Serum cTnT values of 21 patients were below the detection limit (< 0.010 ng/ml). There was no statistical difference between serum cTnT levels of the patients with normal and abnormal echocardiographic findings (P = 0.376). CONCLUSIONS: No correlation was found between serum cTnT values, cumulative doxorubicin doses, and systolic or diastolic cardiac functions. We can conclude that echocardiographic follow-up is more reliable than serum cTnT levels for detecting subclinical cardiac toxicity.