A Nationwide Nepalese Study to Establish Reference Intervals for Major Biochemical Analytes with Elucidation of Nepalese Features of Reference Values.

Despite immense importance of reference intervals (RIs) for clinical diagnosis, there have been no reliable RIs available for Nepalese. Hence, this nationwide study was organized to establish RIs for 30 common biochemical parameters. This study was conducted following the harmonized protocol provided by IFCC Committee on Reference Interval and Decision Limits (C-RIDL) with recruitment of 617 apparently healthy volunteers (18 - 65 years) by near-equal gender balance from 5 major cities. Fasting blood were collected, serum was separated and measured collectively using Beckman-Coulter/Olympus AU480 chemistry analyzer. The sources of variations of reference values (RVs) were evaluated by multiple regression analysis and nested ANOVA. Latent abnormal values exclusion (LAVE) method was applied to reduce influence of latent diseases. RIs were standardized based on a value-assigned serum panel provided by C-RIDL. By ANOVA, no between-city differences were observed, while sex-related changes were typically noted for urate, creatinine, iron, γ-glutamyl transferase (GGT), immunoglobulin M, and transferrin, but not for high-density lipoprotein cholesterol. Age-related changes were observed for total cholesterol (TC), triglyceride, low-density lipoprotein cholesterol, and C-reactive protein (CRP). RIs were successfully derived all parametrically. The LAVE procedure was effective in lowering upper limits for aspartate aminotransferase, alanine aminotransferase (ALT), and CRP. Compared to other collaborating countries, Nepalese RIs were low for urea, cholesterols, ALT, and high for triglyceride, GGT, CRP, immunoglobulin G, and complements. The RIs for major chemistry analytes were derived and standardized for nationwide use in Nepal. This study distinctly elucidated sources of variation and international features of Nepalese RIs. Supplementary Information: The online version supplementary material available at 10.1007/s12291-023-01123-6.

Reference intervals for knee functions specific to outpatients with knee osteoarthritis: a cross-sectional study.

BACKGROUND: Reference values (RVs) for knee function tests have been reported in perioperative patients with knee osteoarthritis (KOA); however, such values for practical use in outpatient setting has yet to be determined. Therefore, we aimed to establish the reference intervals (RIs) for outpatients with mild to moderate KOA. METHODS: This cross-sectional study enrolled 202 outpatients with KOA from 8 Japanese orthopedic clinics and measured knee extensor/flexor muscle strength (MS) and knee extension/flexion range of motion (ROM). We used multiple regression analysis to evaluate the sources of variation, including sex, age, body mass index, Kellgren-Lawrence (K-L) classification, bilateral KOA, and exercise habits. Magnitude of between-subgroup differences is expressed as standard deviation ratio (SDR) based on a three-level nested analysis of variance, with SDR ≥ 0.4 as the threshold for requiring RIs specific for subgroups. RIs were calculated parametrically using two-parameter Box-Cox formula if Gaussian transformation of RVs was successful, otherwise calculated nonparametrically. RESULTS: Partitioning was required by sex for extensor and flexor MS (SDR = 0.65, 0.57, respectively) and by K-L classification for flexion ROM (SDR = 0.54). RIs were determined parametrically for extensor MS as 0.27-2.09 (male) and 0.27-1.54 (female) Nm/kg and for flexor MS 0.18-1.20 (male) and 0.13-0.79 (female) Nm/kg. On the other hand, RIs for extension and flexion ROM were determined nonparametrically due to discrete nature of their RVs. The RIs determined for extension ROM were -15°-0° and for flexion ROM were 105°-150° (for K-L grade I/II) and 95°-140° (for K-L grade III/IV). CONCLUSIONS: The ranges of RIs determined specifically for patients with mild to moderate KOA were in-between those of age-matched healthy controls and pre-surgical KOA patients, both of which we had reported for use in physiotherapeutic management of KOA patients undergone total knee arthroplasty. The newly derived RIs will provide an objective benchmark for physiotherapy targeting outpatients with mild to moderate KOA.

Reappraisal of serum retinol-binding protein as a surrogate marker for retinol and discovery of a novel retinol estimation formula.

BACKGROUND & AIMS: Serum retinol (ROH) is commonly used for population level assessment of vitamin A status. High-performance liquid chromatography (HPLC) is considered most accurate method for measuring ROH. However, with the technical difficulty of using HPLC for routine assays, serum retinol-binding protein (RBP) measured by immunological assays is expected to be a surrogate marker for ROH, with reports of a close correlation between serum RBP and ROH. Nevertheless, RBP is not commonly tested to assess vitamin A status with concerns over RBP alterations under various physiopathological conditions. Thus, we reappraised the extent to which RBP could be used as a surrogate marker in representative disorders that alter serum RBP levels. As a related marker, diagnostic utility of transthyretin (TTR) was also evaluated. METHODS: To evaluate the reliability of ROH and RBP assays, specimen stability was assessed in terms of (1) storage at 25, 4, -20, and -80 °C for 1-28 days, (2) five-cycle freeze-thawing, and (3) fluorescent light exposure for 1-14 days. Sources of variation (sex, age, body mass index [BMI], and drinking habits) and reference intervals for ROH, RBP, and TTR were determined in 617 well-defined healthy individuals. To investigate the influence of disorders that affect serum RBP, patients with five diagnostic groups were enrolled: 26 with chronic kidney disease (CKD); 13 with various malignancies in advanced stages (AdM), 12 with acute bacterial infections (ABI), 6 with liver cirrhosis (LC), and 26 with simple obesity (BMI ≥ 27 kg/m2). RESULTS: The stability of RBP and ROH in serum was confirmed under all conditions. In healthy individuals, serum ROH, RBP, and TTR were appreciably high in males with a slight increase in proportion to age and BMI. The major-axis regression line between RBP (x) and ROH (y) in healthy individuals was y = x, with a correlation coefficient of 0.986. In the LC, AdM, and ABI groups, similar strong correlations were observed; however, the regression lines were shifted slightly rightward from the healthy group line, indicating a positive bias in estimating ROH. Interestingly, the same analyses between TTR and ROH revealed similar strong linear relationships in all groups; however, the regression line of each group showed a leftward (opposite) shift from the healthy group line. Based on these observations, we developed a novel regression model composed of RBP and TTR, which gave much improved accuracy in estimating ROH, even under these pathological conditions. CONCLUSIONS: The perfect RBP-ROH correlation in healthy individuals indicates the utility of RPB as a surrogate marker for ROH. Nevertheless, under RBP-altered conditions, a slight overestimation of ROH is inevitable. However, when the TTR was tested together, the bias can be corrected almost perfectly using the novel ROH estimation formula comprising RBP and TTR.

The Utility of a Novel Stacked Microvascular Imaging for Enhanced Detection of Fibrosis in Chronic Liver Diseases.

OBJECTIVE: Ultrasonographic imaging plays a primary role to detect fibrotic changes in patients with chronic liver disease (CLD). To enhance detectability of fibrosis in its early stage, we developed a novel stacked microvascular imaging (SMVI) that enables continuous visualization of fibrotic changes in intrahepatic vessels. METHODS: SMVI was produced by accumulating 3-5 seconds of high-definition color images in tilted-scan mode. An SMVI score was devised by quantitating three hallmark vascular changes in liver fibrosis in 0-2 grades (total 0-6): narrowing, caliber irregularity, and tortuosity. To evaluate the clinical utility of the SMVI score, 469 well-defined CLD patients were enrolled and subgrouped by the stage of liver fibrosis defined based on elastography: F0-1Low, F0-1High, F2, F3, and F4. The diagnostic performance of the SMVI score was compared to conventional B-mode liver morphology score and various laboratory test markers of fibrosis. RESULTS: Unlike conventional microvascular imaging that relies on a single image, SMVI enabled an undisrupted view of intrahepatic vessels for easy detection of fibrotic changes. SMVI detected microvascular narrowing in 92% at stage F0-1High. While detection rates for caliber irregularity and tortuosity were low at early stages but increased proportionately in advanced stages. Multiple logistic regression analysis revealed that SMVI score was most accurate in distinguishing F0-1Low from F0-1High cases compared to B-mode or laboratory test scores. CONCLUSION: SMVI provides enhanced vascular images of liver fibrosis in CLD, especially in its early stage. The SMVI score can be used as a primary tool for determining fibrotic stages in CLD.

Establishment of reference interval for hemoglobin A1C and other hemoglobin subfractions for healthy Saudi adults.

BACKGROUND: The establishment of Reference Intervals (RIs) for Hemoglobin A1C and other hemoglobin subfractions (A1A, A1B, F, LA1C, A0) is of utmost importance in screening, diagnosing, and monitoring diabetes and other hemoglobin abnormalities through the application of high-pressure liquid chromatography (HPLC) technique. Because there are no locally established RIs for these parameters, it is essential to establish RIs specific to the Saudi population to accurately diagnose and monitor diabetic individuals and identify abnormal levels in hemoglobin subfractions. METHODS: As part of the IFCC global multicenter study of laboratory reference values, a cross-sectional study was conducted in Saudi Arabia. The study involved recruiting a total of 381 healthy adult subjects (>18 years, BMI 28.3 ± 6 kg/m2). Blood samples were analyzed for A1C, biochemical and other immunoassay parameters. The need for RIs based on sex, age, and BMI was determined using the standard deviation ratio (SDR) through a 3-level nested ANOVA. RESULTS: Based on the threshold of SDR≥0.4, RIs for A1C and other Hb subfractions were not partitioned by sex or BMI, but partitioned by age (<45 & ≥45 years) for A1C, LA1C, A0 and F. Spearman's correlation between glucose, insulin, and C-peptide showed a positive association with different hemoglobin subtractions of A1B, F, A1C, and LA1C. The RIs were obtained by using the parametric method and the latent abnormal values exclusion (LAVE) principle was applied on A1C. CONCLUSION: This study established RIs for A1C and other Hb subfractions for healthy adult Saudis. Age was found to be an important source of variation for most of the parameters including A1C. These findings will enhance the understanding and clinical decision-making concerning A1C and other hemoglobin subfractions. The elevated upper limit of RIs for A1C reflects the high prevalence of diabetes in the Saudi population specially in those with increased age.

Establishment of reference intervals for free light chains and immunoglobulins in Saudi population.

OBJECTIVES: Testing of serum-free light chains kappa (κ) and lambda (λ), along with ratio (FLCR) is essential for the diagnosis and management of monoclonal gammopathies. Accurate clinical diagnosis depends upon appropriate local population reference intervals (RIs). This study examined the Saudi population for serum-free light chains and other immunoglobulins to establish RIs and to explore variations in the test results by using the International Federation for Clinical Chemistry and Laboratory Medicine's global protocol for harmonized implementation of RI study. METHODS: A total of 180 healthy Saudi adults were recruited. All serum samples were assayed using the Freelite reagents from the Binding Site. The variation in reference values attributable to sex, age, BMI, and region was calculated by ANOVA as a standard deviation ratio (SDR). The RIs for the FLCR were derived by the parametric method and validated by using samples from patients with hypo- and hypergammaglobulinemia. RESULTS: The new RIs for free κ and FLCR were shifted to a higher side from the manufacturer-adapted RIs. Based on the SDR cutoff value (>0.4), between-sex partition RIs were not required for all analytes except IgM. Validation using patients with hypo- or hypergammaglobulinemia and without multiple myeloma, was all within the new RI. BMI, smoking, and exercise were not relevant sources of variation for any analyte. CONCLUSIONS: Locally derived RIs for free light chains and immunoglobulins analytes specific for Saudis were established after careful consideration of various factors. These RIs were more reliable than those provided as guidance by the manufacturer, or from other countries, for appropriate classification and prediction of disease progression for Saudi patients.

Critical appraisal of two Box-Cox formulae for their utility in determining reference intervals by realistic simulation and extensive real-world data analyses.

BACKGROUND: The reference interval (RI) is defined as the central 95 % range of reference values (RVs) from healthy individuals. The ideal method for determining RIs is to transform RV distribution into Gaussian and estimate its 95 % range parametrically. One-parameter Box-Cox formula (1pBC) is widely used for correcting skewness (Sk) or kurtosis (Kt) in data distribution. However, 1pBC is not popular for computing RIs due to its unreliability in Gaussian transformation. While its two-parameter version (2pBC) is not used due to a challenge in fitting power (λ) and shift (α) parameters simultaneously. In this study, technical issues in fitting both formulae are assessed, and an alternative algorithm for successful use of 2pBC is proposed. METHODS: For fitting 1pBC, optimal λ was determined by stepwise linear search. For 2pBC, optimal [λ, α] combination was pursued in two ways: by grid search of λ and α (2pBCgrid) or by using the grid search but keeping α-range close to the reference distribution (2pBCopt). Their accuracy and precision in determining RIs were compared by generating power-normal distributions simulating RVs of 23 major chemistry analytes. Additionally, their practical utilities were compared by analyzing 776 real-world datasets comprising test results of 25 analytes that were obtained from the global multicenter RV study of IFCC. For comparison, the performance of nonparametric method was evaluated in both settings. RESULTS: For analytes with not-much-skewed distributions, unbiased estimation of RIs was accomplished by all methods. Nevertheless, when reference distributions are located far from zero, λ estimated by1pBC and 2pBCgrid fluctuated widely, which was attributable to virtually flat goodness-of-fit profile for [λ, α]. For highly skewed distributions, 1pBC caused bias in estimating RI and λ due to remotely peaked goodness-of-fit profile. Real-world data analyses revealed that 2pBCopt and 1pBC achieved Gaussian transformation (|Sk|<0.1 and |Kt|<0.3) in 82.4 % and 66.9 % among 776 datasets, respectively. Fitting bias signified by Kt<-0.4 was more common to 1pBC. The practical utility of 2pBCopt was unbiased prediction of analyte-specific distribution-shape (λ). Whereas nonparametric method gave highly variable RIs for both simulated and real-world datasets. CONCLUSIONS: 2pBCopt is suitable for calculating RIs and grasping distribution-shape from the estimated λ.

Reference intervals and sources of variation of pressure pain threshold for quantitative sensory testing in a Japanese population.

Quantitative sensory testing (QST) is useful when analysing musculoskeletal pain disorders. A handheld algometer is most commonly used for pressure pain threshold (PPT) tests. However, reference intervals for PPTs are not elucidated. We assessed reference intervals of PPTs for QST in 158 healthy adult Japanese with no history of musculoskeletal or neurological problems. A handheld algometer was used to record PPT at five different assessment sites on the body: lumbar paravertebral muscle, musculus gluteus maximus, quadriceps, tibialis anterior muscle, and anterior talofibular ligament. Multiple regression analysis was performed to explore sources of variation of PPT according to sex, age, body mass index, UCLA Activity Level Rating, and Tegner Activity Score. Reference intervals were determined parametrically by Gaussian transformation of PPT values using the two-parameter Box-Cox formula. Results of multiple regression analysis revealed that age was significantly associated with PPT of lumbar paravertebral muscle and musculus gluteus maximus. In females, body mass index showed significant positive correlation with PPT of anterior talofibular ligament, and UCLA Activity Level Rating also showed significant positive association with tibialis anterior muscle and anterior talofibular ligament. Site-specific reference intervals of PPTs for Japanese are of practical relevance in fields of pain research using a handheld algometer.

Establishment of reference intervals for hematological parameters of adult population in the western region of Saudi Arabia.

BACKGROUND: Most of hematology laboratories in Saudi Arabia utilize the reference intervals (RIs) provided by instrument manufacturers. This study aimed to define RIs of hematological parameters for adult population in the western region of Saudi Arabia and to explore their specific features from an international perspective. METHOD: This study was conducted according to the harmonized protocol of IFCC Committee on RIs and Decision Limits. Blood samples collected from 409 healthy Saudi males and females adults were analyzed for complete blood count (CBC) by using Cell-Dyn Sapphire analyzer and for iron profile by using Architect analyzers. The needs for RIs partitioned by sex and age was based on standard deviation ratio (SDR) and/or bias ratio (BR). RIs were derived parametrically with/without application of the latent abnormal values exclusion method (LAVE). RESULTS: Based on thresholds of SDR≥0.4 and/or BR≥0.57, RIs were partitioned by sex for red-blood cell count, hemoglobin, hematocrit, red cell distribution width, erythrocyte sedimentation rate, iron, transferrin, ferritin, eosinophil, platelet, plateletcrit, etc. Partitioning by age was not necessary for any of the analytes. LAVE procedure caused appreciable changes in RI limits for most erythrocyte and iron parameters but not for leukocyte parameters. Comparable to other non-IFCC studies on CBC RIs, the RBC and hematocrit (Ht) ranges have shifted to a higher side in both genders. After applying the LAVE method, the male and female RIs for Hb were 4.56 to 6.22 ×106/µL and 3.94 to 5.25 ×106/µL respectively while RIs for Ht were 40.2 to 52.0% and 33.6 to 44.5% respectively. CONCLUSION: LAVE method contributed to reducing the influence of latent anemia in deriving RIs for erythrocyte related parameters. Using the up-to-date methods, the RIs of CBC determined specifically for Saudis will help to improve the interpretation of test results in medical decision making.

Clinical utility of indirect fluorescent assay for IgA class antibodies against Bartonella henselae in serodiagnosis of cat scratch disease in its early stage.

The utility of IgA class antibodies for the serodiagnosis of cat scratch disease (CSD) was evaluated by developing an indirect immunofluorescent assay (IFA) using an antigen obtained by co-cultivating Bartonella henselae ATCC 49882 with Vero cells. Served for evaluation were 101 sera from patients serologically confirmed as CSD with IgG-IFA ≥1:256, and 144 sera from patients clinically suspected of CSD but not serologically confirmed. The sensitivity of the newly developed IgA-IFA in detecting the confirmed cases was 57.4% (58/101), and 75.0% in combination with IgM-IFA. As for the non-confirmed cases, IgA-IFA turned 8.3% cases (12/144) positive, 10 of whom were subsequently diagnosed as CSD of early stage from clinical courses and/or by repeated testing. The 12-case gain was regarded as a significant improvement. Hence, the diagnostic rate of early-stage CSD is expected to be increased by routinely performing IgA-IFA in addition to conventional IgG/IgM-IFA.

Derivation of sex and age-specific reference intervals for clinical chemistry analytes in healthy Ghanaian adults.

OBJECTVIES: This study is aimed at establishing reference intervals (RIs) of 40 chemistry and immunochemistry analytes for Ghanaian adults based on internationally harmonized protocol by IFCC Committee on Reference Intervals and Decision Limits (C-RIDL). METHODS: A total of 501 healthy volunteers aged ≥18 years were recruited from the northern and southern regions of Ghana. Blood samples were analyzed with Beckman-Coulter AU480 and Centaur-XP/Siemen auto-analyzers. Sources of variations of reference values (RVs) were evaluated by multiple regression analysis (MRA). The need for partitioning RVs by sex and age was guided by the SD ratio (SDR). The RI for each analyte was derived using parametric method with application of the latent abnormal values exclusion (LAVE) method. RESULTS: Using SDR≥0.4 as threshold, RVs were partitioned by sex for most enzymes, creatinine, uric acid (UA), bilirubin, immunoglobulin-M. MRA revealed age and body mass index (BMI) as major source of variations of many analytes. LAVE lowered the upper limits of RIs for alanine/aspartate aminotransferase, γ-glutamyl transaminase and lipids. Exclusion of individuals with BMI≥30 further lowered the RIs for lipids and CRP. After standardization based on value-assigned serum panel provided by C-RIDL, Ghanaian RIs were found higher for creatine kinase, amylase, and lower for albumin and urea compared to other collaborating countries. CONCLUSIONS: The LAVE effect on many clinical chemistry RIs supports the need for the secondary exclusion for reliable derivation of RIs. The differences in Ghanaian RIs compared to other countries underscore the importance of country specific-RIs for improved clinical decision making.

Determination of diagnostic threshold in harmonization and comparison of clinical utility for five major antiphospholipid antibody assays used in Japan.

BACKGROUND: Anticardiolipin antibodies (aCL) and anti-ß2 -glycoprotein I antibodies (aß2 GPI) are essential in diagnosing antiphospholipid syndrome (APS) according to the international APS guideline. Five commercial assays for aCL and aß2 GPI are available in Japan, but their test results are quite discordant. For harmonization of diagnosing APS, upper reference limit (URL) and diagnostic accuracy of each assay were evaluated and compared by testing common sets of specimens across all assays. METHODS: We evaluated two manual and three automated assays for aCL and aß2 GPI of IgG- and IgM classes. 99%URL (the upper limit of reference interval: as per guideline) together with 97.5%URL were determined by testing sera from 198 to 400 well-defined healthy subjects. Both URLs were compared with the cutoff values, which were determined based on ROC analysis by testing 50 each of plasma specimens from patients with/without APS. Diagnostic accuracy was evaluated as area under curve (AUC) of the ROC curve. RESULTS: A variable degree of discrepancy between URLs and the cutoff values was observed, which was partly attributable to between-year assay variability. 97.5%URLs were set lower and closer to the cutoff values than 99%URLs. For all assays, diagnostic accuracies of both aß2 GPI-IgG and aCL-IgG were generally high (AUC: 0.84-0.93); whereas those for IgM-class assays were low (AUC: 0.57-0.67), implicating its utility is limited to rare IgG negative APS cases. CONCLUSION: To ensure harmonized APS diagnosis, the diagnostic thresholds of the five assays were evaluated by common procedures. Contrary to the guideline, 97.5%URL is rather recommended for diagnosing APS, which showed a closer match to the cutoff value.

Exploring the seasonal and regional features of cat-scratch disease on the basis of anti-Bartonella henselae IgM/IgG positive rates in Japan.

OBJECTIVES: This study aimed to explore the seasonal and regional features of cat-scratch disease (CSD) based on 15-years of test results for anti-Bartonella henselae IgG and IgM by immunofluorescence assay (IFA) performed as a laboratory specialized in diagnostic testing of CSD in Japan. A literature search was performed to put our findings in perspective. METHODS: A total of 956 sera from patients suspected of CSD were submitted to our laboratory from nationwide. Seasonal changes in the monthly positive rates of IgG/IgM antibodies and regional distribution of the test specimens were analyzed. RESULTS: The monthly positive rates of anti-B. henselae IFA of IgG and IgM were both significantly high between September and January and low between March and July. The seasonal pattern observed in this study was similar to the ones reported from US and France, which were analyzed from a clinical database (monthly incidence of CSD diagnosis) or from monthly positive rates of either B. henselae PCR or anti-B. henselae IFA. However, fluctuations in the IFA monthly positive rates in this study were more pronounced than other reports. Regarding regionality, the test specimens submitted to us for IFA were prominently more from southwestern areas than from northern/middle-northern areas of Japan. The distribution coincided well with the regional distribution of CSD case reports and with a known regional prevalence of Bartonella-species bacteremia among pet cats in Japan. CONCLUSION: These epidemiological features in Japan are of relevance in the clinical diagnoses of CSD.

Nomogram of age-specific anti-Müllerian hormone levels in healthy Egyptian females.

BACKGROUND: Anti-Müllerian hormone (AMH) is an important determinant of ovarian reserve in fertility workups in many clinical settings. Thus, we investigated the age dependent decline in AMH specific to the Egyptian population and sought to establish an age dependent reference interval parametrically. METHODS: Serum samples were collected from 841 apparently healthy women. AMH was measured using an electro-chemiluminescent technique. Box-Cox power transformation was used to make the AMH distribution Gaussian for parametric derivation of reference intervals. RESULTS: Power of 0.4 was found optimal for Gaussian transformation of AMH reference values. We demonstrate the strong negative relation between circulating AMH and female age with Spearman's correlation coefficient of rS = -0.528. Age-specific reference interval was determined for every 5 years of age from 16 to 49, and nomogram was constructed by smoothing the lines connecting adjacent lower and upper reference limits. CONCLUSION: The age-specific reference intervals and the age-AMH nomogram could be valuable in the clinical practice of in reproductive medicine. To our knowledge, this is the first study to confirm AMH levels in Egyptian females. We were able to explore age-related AMH levels specific to Egyptian females in the fertile age group and to treat skewed AMH data in a multi-step scheme using power transformation. Thus, a more accurate nomogram was constructed accommodating a profile delineated for a wide age range and a rescaled AMH axis improving its usability.

Comparison of reference intervals derived by direct and indirect methods based on compatible datasets obtained in Turkey.

BACKGROUND: Indirect derivation of reference intervals (RIs) from the laboratory information system (LIS) has been recently pursued. We aimed at evaluating the accuracy of indirectly predicted RIs compared to the RIs established directly from healthy subjects in the nationwide RI study in Turkey, targeting 25 major chemistry analytes. METHODS: LIS data were retrieved from the laboratory that performed measurements for the direct study. They were cleaned by limiting to outpatients with age 18-65 years, and by allowing only one record per year per patient. Evaluated were four indirect methods of univariate approach: Hoffmann, Bhattacharya, Arzideh, and Wosniok methods. Power transformation of the LIS dataset was performed either using the power (λ) reported by the IFCC global RI study (the first two methods) or using a λ predicted (the last two). RESULTS: Compared to the direct study dataset, the LIS dataset showed a variable degree of alterations in peak location and shape. Consequently, lower-side peak-shifts observed in sodium, albumin, etc. led to lowered RI limits, whereas higher-side peak-shift observed in triglyceride, low-density lipoprotein cholesterol, etc. led to raised RI limits. Overall, 72% (62-81) of the RI limits predicted by indirect methods showed significant biases from direct RIs. However, the biases observed in total cholesterol, lactic dehydrogenase, etc. were attributed to a higher-side age-bias in LIS dataset. After excluding them, the overall proportion of biased RIs was reduced to 47% (38-54). CONCLUSION: To reduce prediction biases that remained after age adjustment, it is necessary to apply more rigorous data-cleaning before applying indirect methods.

Exploration and time-serial validation of logistic regression models composed of multiple laboratory tests for early detection of HCV-associated hepatocellular carcinoma.

BACKGROUND: We developed a laboratory test-based regression model for early detection of hepatocellular carcinoma (HCC) associated with HCV in its surveillance. METHODS: This matched case-control study was conducted by enrolling 452 patients with chronic hepatitis and/or cirrhosis, including 129 patients complicated with HCC. One-to-one propensity score matching was performed by referring to sex, age, and fibrosis-4 index, which resulted in 102 patients each in HCC and non-HCC groups. Logistic regression models (LRM) for distinguishing the two groups were explored from variable combinations of laboratory tests. The model was validated by our new scheme of applying it retroactively to trimonthly previous datasets. RESULTS: Models with a practical level of diagnostic accuracy (C-statistic) were α-fetoprotein (AFP) alone (0.810), LRM3 comprising AFP, AST, and ALT (0.850), and LRM4 comprising AFP, AFP/(AST × ALT), and AST (0.862). After retroactive application of each model, LRM4 showed the highest distinction of the two groups at -12M, -6M, -3M with C-statistics of 0.654, 0.786, 0.834, respectively. LRM4 was accurate even after limiting cases to early-stage HCC. CONCLUSIONS: LRM4 was proved useful in prompting clinicians to perform timely image study in the surveillance. The retroactive validation scheme is applicable to assess diagnostic models of other neoplastic diseases.

Factors characterizing gait performance of patients before and soon after knee arthroplasty.

[Purpose] For monitoring patients with knee osteoarthritis undergoing knee arthroplasty, the Timed Up and Go and maximum walking speed tests are commonly used. To provide appropriate peri-surgical rehabilitation, we evaluated the factors associated with postsurgical changes in Timed Up and Go and maximum walking speed results. [Participants and Methods] We enrolled 545 knee osteoarthritis patients undergoing either of the following knee arthroplasties: conventional total knee arthroplasty, minimally invasive total knee arthroplasty, and unicompartmental knee arthroplasty. Comfortable Timed Up and Go, maximum Timed Up and Go, and maximum walking speed were measured 2 weeks before and soon after surgery. Factors (gender, age, and surgical mode) that might influence changes in test results were evaluated by multiple regression analysis and a two-factor stratification diagram. [Results] Multiple regression analysis revealed that postsurgical changes in comfortable/maximum Timed Up and Go and maximum walking speed results were associated with age and surgical mode after adjustment for preoperative values. Two-factor diagrams showed that the older the patient, the greater was the slowdown in the Timed Up and Go test performed postoperatively. The levels of slowdown in the postoperative Timed Up and Go and maximum walking speed tests were the smallest in those who underwent conventional total knee arthroplasty, followed by those who underwent minimally invasive and unicompartmental knee arthroplasty. Among patients whose preoperative Timed Up and Go and maximum walking speed were slow, slowdown in Timed Up and Go was pronounced with age, and slowdown in maximum walking speed was higher in conventional total knee arthroplasty. [Conclusion] The changes in Timed Up and Go and maximum walking speed results 2 weeks after knee arthroplasty depended on age and surgical modes. These findings are relevant for the implementation of appropriate peri-surgical rehabilitation.

Determination of reference intervals for knee motor functions specific to patients undergoing knee arthroplasty.

BACKGROUND: In patients with knee osteoarthritis (KOA) undergoing knee arthroplasty (KA), lower-limb motor function tests are commonly measured during peri-surgical rehabilitation. To clarify their sources of variation and determine reference intervals (RIs), a multicenter study was performed in Japan. METHODS: We enrolled 545 KOA patients (127 men; 418 women; mean age 74.2 years) who underwent KA and followed a normal recovery course. The surgical modes included total KA (TKA), minimally invasive TKA (MIS-TKA), and unicompartmental KA (UKA). Motor functions measured twice before and two weeks after surgery included timed up-and-go (TUG), maximum walking speed (MWS), extensor and flexor muscle strength (MS), and knee range of motion (ROM). Multiple regression analysis was performed to evaluate their sources of variation including sex, age, BMI, and surgical mode. Magnitude of between-subgroup differences was expressed as SD ratio (SDR) based on 3-level nested ANOVA. SDR≥0.4 was set as the threshold for requiring RIs specific for each subgroup. RESULTS: Before surgery, age-related changes exceeding the threshold were observed for TUG and MWS. Between-sex difference was noted for extensor and flexor MS, but extension and flexion ROMs were not influenced by sex or age. After surgery, in addition to similar influences of sex and age on test results, surgical modes of UKA and MIS-TKA generally had a favorable influence on MWS, extensor MS, and flexion ROM. All motor function test results showed a variable degree of skewness in distribution, and thus RIs were basically derived by the parametric method after Gaussian transformation of test results. CONCLUSIONS: This is the first study to determine RIs for knee motor functions specific to KOA patients after careful consideration of their sources of variation and distribution shapes. These RIs facilitate objective implementation of peri-surgical rehabilitation and allow detection of patients who deviate from the normal course of recovery.

Biological sources of variations of tartrate-resistant acid phosphatase 5b in a healthy Japanese population.

BACKGROUND: Tartrate-resistant acid phosphatase 5b (TRACP5b) is a bone resorption marker that is mainly used in clinical management of osteoporosis. For proper interpretations of test results for serum TRACP5b, we explored their biological sources of variation, esp. age-related changes, and associations with other bone-related markers in healthy Japanese adults. METHODS: During the 2009 East-Southeast Asian multicentre study for determination of reference intervals, 72 major laboratory tests were measured by centralized assays in 3541 well-defined healthy volunteers. The current study included 1980 test results in Japanese subjects for five bone-related markers: TRACP5b, bone alkaline phosphatase, intact parathyroid hormone, calcium and inorganic phosphate. Information on sources of variation, including body mass index, smoking habits and ABO-blood group, were obtained from a health status questionnaire. RESULTS: Gender-specific profiles of age-related changes were observed for each parameter. Increased values starting from 40 years of age in females were most prominent for TRACP5b, followed by bone alkaline phosphatase and inorganic phosphate. TRACP5b in males decreased with body mass index, bone alkaline phosphatase and TRACP5b were higher in blood type-O subjects, especially in males. TRACPT5b was closely correlated with bone alkaline phosphatase, and moderately correlated with adjusted calcium and inorganic phosphate, especially in females aged ≥45 years. Reference intervals for each analyte were determined parametrically based on gender and age. CONCLUSIONS: This study elucidated sources of variation of TRACP5b and related bone markers in healthy Japanese subjects and demonstrated a specific age profile for each marker. These results are of relevance for better clinical usage and interpretations of serum levels of bone markers.

Establishment of reference intervals of clinical chemistry analytes for the adult population in Egypt.

BACKGROUND: This is the first Egyptian nationwide study for derivation of reference intervals (RIs) for 34 major chemistry analytes. It was conducted as a part of the global initiative by the IFCC Committee on Reference Intervals and Decision Limits (C-RIDL) for establishing country-specific RIs based on a harmonized protocol. METHODS: 691 apparently healthy volunteers aged ≥18 years were recruited from multiple regions in Egypt. Serum specimens were analyzed in two centers. The harmonization and standardization of test results were achieved by measuring value-assigned serum panel provided by C-RIDL. The RIs were calculated by parametric method. Sources of variation of reference values (RVs) were evaluated by multiple regression analysis. The need for partitioning by sex, age, and region was judged primarily by standard deviation ratio (SDR). RESULTS: Gender-specific RIs were required for six analytes including total bilirubin (TBil), aspartate and alanine aminotransferase (AST, ALT). Seven analytes required age-partitioning including glucose and low-density lipoprotein cholesterol (LDL-C). Regional differences were observed between northern and southern Egypt for direct bilirubin, glucose, and high-density-lipoprotein cholesterol (HDL-C) with all their RVs lower in southern Egypt. Compared with other collaborating countries, the features of Egyptian RVs were lower HDL-C and TBil and higher TG and C-reactive protein. In addition, BMI showed weak association with most of nutritional markers. These features were shared with two other Middle Eastern countries: Saudi Arabia and Turkey. CONCLUSION: The standardized RIs established by this study can be used as common Egyptian RI, except for a few analytes that showed regional differences. Despite high prevalence of obesity among Egyptians, their RVs of nutritional markers are less sensitive to increased BMI, compared to other collaborating countries.