Quality and quantity of visceral fat tissue are associated with insulin resistance and survival outcomes after chemotherapy in patients with breast cancer.

PURPOSE: Recent studies suggest that the quality and quantity of visceral adipose tissue (VAT) play significant roles in adipocyte function, and are related to insulin resistance. We tested the hypothesis that high amounts of upper VAT (aVAT) and low-quality VAT worsen treatment outcomes via altered insulin metabolism. METHODS: Cohort 1 included 106 women with breast cancer who were undergoing surgery. Homeostasis model assessment of insulin resistance (HOMA-R), insulin-like growth factor (IGF)-1, and IGF-binding protein 3 (IGFBP3) were measured before the initiation of treatment. aVAT was measured via computed tomography (CT). VAT quality was assessed using CT-determined Hounsfield units (VAT-HU). Associations between the variables investigated and VAT quality and quantity were analyzed. Cohort 2 included 271 patients who underwent chemotherapy. Associations between the variables investigated and survival outcomes after chemotherapy were analyzed via retrospective chart review. RESULTS: In cohort 1, aVAT was significantly correlated with insulin and HOMA-R levels. As body mass index (BMI) class increased, mean IGF-1 increased and mean IGFBP3 decreased, but these trends were not statistically significant. In cohort 2, aVAT was significantly positively correlated with BMI. The patients in the third aVAT tertiles had significantly shorter distant disease-free survival (dDFS) after neoadjuvant chemotherapy setting. In multivariate analysis, aVAT and VAT-HU were significantly associated with shorter dDFS. CONCLUSIONS: High aVAT and low-quality VAT were associated with poor survival outcome, increased insulin levels, and insulin resistance. The present study suggests the importance of evaluating the quality and quantity of VAT when estimating insulin resistance and treatment outcomes.

Change in albumin measurement method affects diagnosis of nephrotic syndrome.

BACKGROUND: Although serum albumin levels (sALB) have been quantified by the dye-binding method with bro- mocresol green (BCG) or bromocresol purple (BCP) on a routine basis, accurate measurement of sALB with these methods is difficult. The modified BCP method is highly specific to albumin without being affected by sample preservation to enable stable and accurate quantification of albumin concentrations. A change in the albumin measurement method may alter the diagnosis of nephrotic syndrome. METHODS: sALB was measured in 295 patients including 98 patients with chronic renal disease by the modified BCP method, BCG method, and immunonephelometry as the gold standard. RESULTS: sALB measured by the modified BCP method was well correlated with levels measured by immunonephelometry. sALB obtained by the BCG method was significantly higher than the levels measured by the modified BCP method (p < 0.001, Student's t-test). This tendency was more evident in patients with chronic renal disease than other patients. When the threshold value of sALB for the diagnosis criteria of nephrotic syndrome (≤ 25 g/L) and a high risk of thrombosis (≤ 20 g/L) in nephrotic syndrome was based on the BCG method, the revised criteria in the modified BCP method would be ≤ 20.5 and ≤ 14.9 g/L, respectively. CONCLUSIONS: Overestimation of sALB by the BCG method affected diagnosis of nephrotic syndrome. The method by which sALB is measured should be specified in both clinical and research settings in nephrology.

[Usefulness of the standardized serum cystatin C reagent in evaluation of renal function].

Serum cystatin C (sCysC) is valuable to evaluate the renal functions in various clinically studies. Since the reagents to detect sCysC are different among institutes or hospitals, the accidental error due to the difference of reagents is not negligible. That is why sCysC is sometimes difficult to use or interpret for patient as estimated Glomerular filtration rate (eGFR) that is one of the most popular factors to evaluate renal function. Recently, the international reference standard is gradually accepted as standardization of chemical measurement of sCysC in Japan. Japan Society of Clinical Chemistry has reported the utilities for sCysC measurement standardization in 2010. In this study, we examined the usefulness of the standardization of sCysC reagent in Chiba University Hospital. Our study indicated that the clinical usefulness of eGFR calculated by sCysC. As results, eGFR from serum creatinine (sCr) was relatively high in patients with reduced muscle mass, such as old-age patients with wheelchair and prolonged hospitalization. On the other hand, eGFR from sCysC was high in patients with hypothyroidism. Together, eGFR calculated by sCysC is clinically available for the patients with reduced muscle mass could be underestimated. Further studies are required to evaluate the validity of standardization of sCysC measurement and find the dissociation among eGFR, sCysC and sCr depending in the renal disease or pathogenesis.

Serum cystatin C as a marker for early detection of chronic kidney disease and grade 2 nephropathy in Japanese patients with type 2 diabetes.

BACKGROUND: Diabetic nephropathy (DN) is a significant cause of hemodialysis, and its early detection is extremely important to prevent or delay end-stage renal disease. The significance of the renal function marker serum cystatin C (sCysC) and its relationship with glomerular filtration rate in chronic kidney disease (CKD) and DN in Japanese patients with type 2 diabetes remains uncertain. In this study, we examined the effectiveness of sCysC as a marker of early DN and CKD in Japanese subjects. METHODS: A total of 325 Japanese patients with type 2 diabetes and 88 healthy subjects were studied retrospectively. sCysC concentration (mg/L) was determined by a latex turbidmetric immunoassay using a BioMajesty 8040 analyzer. The renal function of the diabetic patients was evaluated using the albumin-creatinine ratio (ACR) and Kidney Disease Outcome Quality Initiative-Kidney Disease Improving Global Outcomes (K/DOQI-KDIGO) classification. RESULTS: There was a significant increase in sCysC but not in serum creatinine (sCr) or serum ß2-microglobulin (sß2M) in patients with grade 2 DN (ACR 30-300 mg/g) compared to grade 1 patients. Receiver operating characteristic analysis in grade 2 and 3 DN patients showed that sCysC had superior sensitivity and specificity than sCr and sß2M for early detection of DN. In addition, sCysC showed particularly high sensitivity and specificity in DN patients with stage 2 CKD. CONCLUSIONS: sCysC was effective for detection of grade 2 DN and would be especially useful for screening stage 2 CKD patients (K/DOQI-KDIGO).

[Clinical validity of renal function markers including serum cystatin C on chronic kidney disease classification].

In this study, clinical utility of various renal function markers (Cystatin C, Creatinine, beta2-microglobulin, Urea nitrogen) including serum Cystatin C was evaluated by the latex turbidimetric immunoassay based on the Chronic Kidney Disease (CKD) stage classification. Serum creatinine was most correlated with estimated Glomerular Filtration Rate (eGFR), followed by serum Cystatin C. However, the level of serum Cystatin C increased significantly earlier than other renal function markers in the group of mild renal dysfunction based on the CKD stage classification. Cystatin C also had the most distinctive ability of the renal dysfunction by Receiver Operating Characteristic (ROC) and distinction characteristic analysis. Thus, it is useful to measure serum Cystatin C and serum creatinine at the same time to improve diagnostic sensitivity and specificity, eGFR was 6% higher than Ccr on stage III-V, though Ccr was 30% higher than eGFR on stage I-II by CKD stage classifications. Since eGFR and Ccr showed different results in early and severe CKD groups, it was suggested that Ccr and eGFR need to be evaluated as different renal function markers.

Reducing the incidence of pseudohyperkalemia by avoiding making a fist during phlebotomy: a quality improvement report.

BACKGROUND: Pseudohyperkalemia is uncommon, but important. Local release of potassium caused by contraction of the forearm muscles from a tightly clenched fist or repeated fist clenching during phlebotomy is a recognized cause of pseudohyperkalemia. We investigated the use of a standard protocol to avoid fist clenching during phlebotomy. STUDY DESIGN: Quality improvement report. SETTING & PARTICIPANTS: In 7 healthy volunteers, 10 blood samples were collected over 10-second intervals after 20 repeated fist clenching and unclenching movements. In 86 healthy volunteers, 3 blood samples were collected with and without prior fist clenching. Between September 1, 2006, and June 30, 2007, peripheral venous blood samples were collected from 73,846 outpatients at Chiba University Hospital without a protocol to avoid fist clenching. Between July 1, 2007, and March 31, 2009, blood samples were collected from 171,053 outpatients using the protocol. QUALITY IMPROVEMENT PLAN: After July 1, 2007, blood samples were collected from the basilic or cephalic vein without making a fist or by making a fist using minimal gripping strength. Also, when multiple specimens were obtained from 1 patient, the specimen for measuring serum electrolytes was obtained after the other specimens. OUTCOMES & MEASUREMENTS: Pseudohyperkalemia, defined as unexplained serum potassium level ≥6.5 mmol/L. RESULTS: In the 7 volunteers, the decrease in serum potassium levels after cessation of fist clenching ranged from 8.4%-25.9%. In the 86 volunteers, the percentage with a decrease in serum potassium level ≥0.2 mmol/L between the first and third samples was 25.6% versus 6.7% with or without prior fist clenching, respectively. In clinical practice, we observed 8 cases of pseudohyperkalemia before implementing the protocol (0.0081%) and 1 case (0.00058%) after implementing the protocol (P = 0.001). LIMITATIONS: Causes of hyperkalemia before using precautions were assessed using retrospective analyses. CONCLUSIONS: Avoiding fist clenching during phlebotomy and not using the first specimen for electrolyte measurements when obtaining multiple specimens from a single patient can reduce the occurrence of pseudohyperkalemia.

Central blood pressure and pulse wave velocity in young and middle-aged Japanese adults with isolated systolic hypertension.

Isolated systolic hypertension (ISH), defined as systolic blood pressure (SBP) ≥140 mmHg and diastolic BP (DBP) <90 mmHg, is a common type of hypertension among young men. This study aimed to investigate the clinical characteristics, central blood pressure, and arterial stiffness of young and middle-aged Japanese individuals with ISH. A total of 432 male participants, aged 18-49 years, were classified into six subgroups: optimal BP (SBP <120 mmHg and DBP <80 mmHg), high-normal BP (SBP 120-129 mmHg and DBP <80 mmHg), high-BP (SBP 130-139 mmHg and/or DBP 80-89 mmHg), ISH (SBP ≥140 mmHg and DBP <90 mmHg), isolated diastolic hypertension (IDH) (SBP <140 mmHg and DBP ≥90 mmHg), and systolic and diastolic hypertension (SDH) (SBP ≥140 mmHg and DBP ≥90 mmHg). Participants with ISH had a greater body mass index (BMI) and waist circumference than the optimal BP participants but were more likely to be physically active than the IDH and SDH participants. The central SBP of the ISH subgroup was higher than that of the optimal/high-normal/high-BP subgroups and lower than that of the SDH subgroup. The carotid-femoral pulse wave velocity (cfPWV) of the ISH subgroup was higher than that of the optimal and high-normal BP subgroups and lower than that of the SDH subgroup after adjusting for age, heart rate, BMI, and physical activity. These differences disappeared after further adjustment for central mean arterial pressure. In conclusion, the central SBP of Japanese men with ISH was greater than that of Japanese men with optimal/high-normal/high-BP, but the progression of arterial stiffness was unclear.

Development of a simple indocyanine green measurement method using an automated biochemical analyser.

Background The indocyanine green retention rate is important for assessing the severity of liver disorders. In the conventional method, blood needs to be collected twice. In the present study, we developed an automated indocyanine green method that does not require blood sampling before intravenous indocyanine green injections and is applicable to an automated biochemical analyser. Methods The serum samples of 471 patients collected before and after intravenous indocyanine green injections and submitted to the clinical laboratory of our hospital were used as samples. The standard procedure established by the Japan Society of Hepatology was used as the standard method. In the automated indocyanine green method, serum collected after an intravenous indocyanine green injection was mixed with the saline reagent containing a surfactant, and the indocyanine green concentration was measured at a dominant wavelength of 805 nm and a complementary wavelength of 884 nm. Results The coefficient of variations of the within- and between-run reproducibilities of this method were 2% or lower, and dilution linearity passing the origin was noted up to 10 mg/L indocyanine green. The reagent was stable for four weeks or longer. Haemoglobin, bilirubin and chyle had no impact on the results obtained. The correlation coefficient between the standard method (x) and this method (y) was r=0.995; however, slight divergence was noted in turbid samples. Conclusion Divergence in turbid samples may have corresponded to false negativity with the standard procedure. Our method may be highly practical because blood sampling before indocyanine green loading is unnecessary and measurements are simple.

Identification of specific and common diagnostic antibody markers for gastrointestinal cancers by SEREX screening using testis cDNA phage library.

The present study was planned to identify novel serum antibody markers for digestive organ cancers. We have used screening by phage expression cloning and identified novel fourteen antigens in this experiment. The presence of auto-antibodies against these antigens in serum specimens was confirmed by western blotting. As for auto-antibodies against fourteen antigens, AlphaLISA (amplified luminescence proximity homogeneous assay) assay was performed in the sera of gastrointestinal cancers patients to confirm the results. Serum antibody levels against these fourteen recombinant proteins as antigens between healthy donors (HD) and esophageal squamous cell carcinoma (ESCC) patients, gastric cancer (GC), or colon cancer (CC) were compared. The serum levels of all fourteen auto-antibodies were significantly higher in ESCC and GC than those of HD. Among those auto-antibodies, except ECSA2 and CCNL2, were also detected significantly higher levels in CC than those of HD. Receiver operating curve (ROC) revealed similar results except CCNL2 in CC. AUC values calculated by ROC were higher than 0.7 in auto-antibodies against TPI1, HOOK2, PUF60, PRDX4, HS3ST1, TUBA1B, TACSTD2, AKR1C3, BAMBI, DCAF15 in ESCC, auto-antibodies against TPI1, HOOK2, PUF60, PRDX4, TACSTD2, AKR1C3, BAMBI, DCAF15 in GC, and auto-antibodies against TPI1, HOOK2, PUF60 in CC. AUC of the combination of HOOK2 and anti-p53 antibodies in ESCC was observed to be as high as 0.8228. Higher serum antibody levels against ten antigens could be potential diagnostic tool for ESCC. Higher serum antibody levels against eight antigens could be potential diagnostic tool for GC, and serum antibody levels against three antigens could be potential diagnostic tool for CC.

Accelerated oligosaccharide absorption and altered serum metabolites during oral glucose tolerance test in young Japanese with impaired glucose tolerance.

AIMS/INTRODUCTION: Impaired glucose tolerance (IGT) is a subtype of prediabetes, a condition having high risk for development to diabetes mellitus, but its pathophysiology is not fully understood. In the present study, we examined metabolic changes in IGT by using two types (D-glucose [Glc] and partial hydrolysate of starch [PHS]) of oral glucose tolerance tests (OGTTs), with emphasis on serum incretins and metabolites. MATERIALS AND METHODS: We carried out the two types of OGTT (Glc/OGTT and PHS/OGTT) in 99 young Japanese individuals who had tested either positive (GU+ ; n = 48) or negative (GU- ; n = 51) for glycosuria. After OGTT, they were sub-grouped into five categories: normal glucose tolerance (NGT) in the GU- group (GU- /NGT; n = 49), NGT in the GU+ group (GU+ /NGT; n = 28), IGT (n = 12), diabetes mellitus (n = 1) and renal glycosuria (n = 9). Serum incretin and metabolites of GU- /NGT and IGT were then measured. RESULTS: When the serum insulin level at each time-point during PHS/OGTT was expressed as its ratio relative to Glc/OGTT, it was increased time-dependently in GU- /NGT, but not in IGT. Such an increase in the ratio was also detected of serum incretin levels in GU- /NGT, but not in IGT, suggesting a lack of deceleration of oligosaccharide absorption in IGT. Metabolome analysis showed a difference in the serum levels of two metabolites of unknown function in mammals, methylcysteine and sedoheptulose 1,7-bisphosphate, between GU- /NGT and IGT. CONCLUSIONS: Comparison of PHS/OGTT and Glc/OGTT showed that oligosaccharide absorption was accelerated in IGT. Methylcysteine and sedoheptulose 1,7-bisphosphate could be novel markers for dysregulated glucose metabolism.

Paradoxical effects of thyroid function on glomerular filtration rate estimated from serum creatinine or standardized cystatin C in patients with Japanese Graves' disease.

BACKGROUND: Estimated glomerular filtration rate (eGFR) is clinically valuable for evaluating renal function. Recently, serum cystatin C (sCysC) measurement has been standardized and has demonstrated utility as a novel indicator of renal function. Thyroid hormone is known to affect serum creatinine (sCr) and sCysC, however, the clinical significance of post-treatment renal function evaluation is yet to be completely elucidated. This study examined the effects of thyroid hormones on eGFR by sCr (eGFRCr), and standardized sCysC (eGFRCysC) in patients with Japanese Graves' disease (GD). METHODS: Serum samples were obtained from 113 outpatients with GD. Following pharmacotherapy, 41 of the 113 outpatients with GD achieved remission. Renal function was evaluated by eGFRCr and eGFRCysC. Reference method used Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equations. RESULTS: eGFRCr levels significantly increased whereas eGFRCysC levels significantly decreased with elevated FT3 and FT4 levels in patients with GD. In the remission group, eGFRCr levels significantly decreased and eGFRCysC levels significantly increased. No significant differences between eGFRCr and eGFRCysC levels were observed. Furthermore, CKD-EPI equations show a similar trend and eGFRCr-CysC levels were no significant differences regardless of before and after treatment. CONCLUSIONS: Renal function evaluation by eGFRCr and eGFRCysC had clinical utility in post-treatment euthyroidism.

The abnormal reaction data-detecting function of the automated biochemical analyzer was useful to prevent erroneous total-bilirubin measurement and to identify monoclonal proteins.

BACKGROUND: Recently, to detect abnormal reactions and failures of the device in biological analysis, a reaction data monitoring system has been provided for automated biochemical analyzers. We investigated the usefulness of this function for total-bilirubin (T-Bil) measurement in routine testing. METHODS: Abnormal reactions of T-Bil were detected in the reaction data over time based on the following items: whether the absorbance variance after mixing of the first reagent and sample exceeds the cut-off value. RESULTS: In the cases in which the abnormal reaction was observed, the absorbance rapidly rose because of turbidity after mixing the sample with Reagent-1. The measured value was higher than the actual T-Bil level, for which the analyzer showed a warning mark with the output data. When this particular serum sample was subjected to immunofixation electrophoresis, the presence of a monoclonal protein was confirmed. We encountered seven similar cases out of 30,731 samples. CONCLUSIONS: The reaction data monitoring system of the automated biochemical analyzers was useful to prevent false reports (misdiagnosis) due to unpredictable problems during T-Bil measurement. It was also suggested that detection of false reaction with a reagent may be a clue to find a new pathology, such as monoclonal gammopathy.

Estimated glomerular filtration rate by serum creatinine or standardized cystatin C in Japanese patients with Graves׳ disease.

Glomerular filtration rate (eGFR) by serum creatinine (eGFRCr) or standardized cystatin C (eGFRCysC) were estimated in Japanese patients with Graves׳ disease (GD) of different sex. Clinical samples were collected from patients with GD with normal renal function to accurately validate eGFRCr and eGFRCysC levels and evaluate how hyperthyroidism affects renal function. Levels of eGFRCr and eGFRCysC showed clinical usefulness in successfully treated euthyroid patients with GD regardless of sex. The article includes detailed experimental methods and data used in our analysis. The data relates to the "Paradoxical effect of thyroid function on the estimated glomerular filtration rate by serum creatinine or standardized cystatin C in Japanese Graves' disease patients" (Suzuki et al., 2015) [1].

Decreases in the serum VLDL-TG/non-VLDL-TG ratio from early stages of chronic hepatitis C: alterations in TG-rich lipoprotein levels.

BACKGROUND: The liver secretes very-low-density lipoproteins (VLDLs) and plays a key role in lipid metabolism. Plasma total triglyceride (TG) level variations have been studied in patients with hepatitis C virus (HCV)-related chronic hepatitis (CH-C). However, the results of these studies are variable. A homogenous assay protocol was recently proposed to directly measure the TG content in VLDL (VLDL-TG) and VLDL remnants. METHODOLOGY/PRINCIPAL FINDINGS: Using the assay protocol, we determined serum VLDL-TG levels in 69 fasting patients with biopsy-proven HCV-related chronic liver disease and 50 healthy subjects. Patients were classified into stages F0-F4 using the 5-point Desmet scale. Serum total TG levels in patients with non-cirrhotic (F1-F3) CH-C did not demonstrate significant differences compared with healthy subjects, but serum VLDL-TG levels did demonstrate significant differences. Mean serum VLDL-TG levels tended to decrease with disease progression from F1 to F4 (cirrhosis). Compared with healthy subjects, serum non-VLDL-TG levels significantly increased in patients with stages F2 and F3 CH-C; however, we observed no significant difference in patients with liver cirrhosis. Furthermore, the serum VLDL-TG/non-VLDL-TG ratio, when taken, demonstrated a significant decrease in patients with CH-C from the mildest stage F1 onward. CONCLUSIONS/SIGNIFICANCE: The decrease in serum VLDL-TG levels was attenuated by increase in non-VLDL-TG levels in patients with non-cirrhotic CH-C, resulting in comparable total TG levels. Results of previous studies though variable, were confirmed to have a logical basis. The decrease in the serum VLDL-TG/non-VLDL-TG ratio as early as stage F1 demonstrated TG metabolic alterations in early stages of CH-C for the first time. The involvement of TG metabolism in CH-C pathogenesis has been established in experimental animals, while conventional TG measurements are generally considered as poor indicators of CH-C progression in clinical practice. The serum VLDL-TG/non-VLDL-TG ratio, which focuses on TG metabolic alterations, may be an early indicator of CH-C.