Use of Ibrutinib in 10 Patients with Treatment-Naïve or Relapsed/Refractory Chronic Lymphocytic Leukemia/Small Lymphocytic Lymphoma in Real-World Clinical Practice -A Report from a Single Medical Institution.

In Japan, ibrutinib has been approved as both a front-line and later-line treatment for chronic leukemia/small lymphocytic lymphoma(CLL/SLL). However, little is known about the actual outcomes and adverse events(AEs)associated with the use of ibrutinib in Japanese patients. OBJECTIVE: The outcomes and AEs of patients treated with ibrutinib in a real-world setting were investigated. METHODS: A retrospective cohort study of all patients with CLL/SLL who were treated with ibrutinib at a single institution was conducted. RESULT: In total, 10 patients, including 5 treatment-naïve patients(50%), were enrolled. The median follow-up period was 9.8 months(range, 0.2-21.6 months), and the estimated overall response rate (ORR: complete remission plus partial remission)was 60%. The median overall survival and progression-free survival outcomes were not reached. During the follow-up period, 4 patients(40%)had at least one AE and 1 patient(10%)had at least one grade≥3 AE. Ibrutinib was discontinued in 4 patients(40%)because of AEs in 2 patients(20%), the progression of CLL in 1 patient(10%), and financial reasons in 1 patient(10%). Richter's transformation did not occur in any of the cases. CONCLUSION: The ORR was lower(60%)than that observed in clinical trials. The frequency and severity of AEs were both relatively low, although the discontinuation rate was high(40%). Patient education and medication adherence were considered important.

Low‒Risk Essential Thrombocythemia Who Presented with Recurrent Episodes of Cerebral Hemorrhage during Pregnancy and Developed Cerebral Infarction during Puerperium.

A 42‒year‒old woman. At week 27 of pregnancy, she developed subcortical hemorrhage and underwent open cranial surgery for hematoma evacuation. The platelet(Plt)count was 297,000/µL. At week 34 of pregnancy, she developed subcortical hemorrhage again. The Plt count was 429,000/µL. At week 35 of pregnancy, the ventricular rupture and she underwent drainage and emergency cesarean section. The Plt count was 687,000/µL. Two days after delivery, hemorrhage was detected. The Plt count was 815,000/µL. Six days after delivery, she developed infarction. The Plt count was 915,000/µL. MRI revealed no evidence of aneurysm, arteriovenous malformations or tumor. Ten days after delivery, the Plt count was 1,173,000/µL. Bone marrow examination led to the diagnosis of essential thrombocythemia(ET). JAK2, CARL and MPL was negative. She was rated as"low‒risk"by IPSET‒thrombosis, and as"ultralow"risk by revised IPSET‒thrombosis. von Willebrand factor(VWF)activity was as high as 247%. The bleeding time and platelet aggregation activity were normal. There was no evidence of disseminated intravascular coagulation(DIC)or hypertensive disorders of pregnancy(HDP). She died of cerebral hemorrhage and infarction, 26 days after delivery.

Accuracy study of a novel alternate method measuring erythrocyte sedimentation rate for prototype hematology analyzer Celltac α.

INTRODUCTION: The erythrocyte sedimentation rate (ESR) is a nonspecific inflammation indicator. In laboratory testing, automated ESR analyzers may use the reference Westergren method (Reference WG), modified Westergren (Modified WG), or Alternate ESR method (Alternate ESR) based on photometric rheology. A prototype hematology analyzer Celltac α+ (Nihon Kohden Corporation) with built-in Novel ESR analysis technology (Novel ESR) was developed to improve the accuracy of Alternate ESR. Alternate ESR uses only the aggregation phase information of Reference WG. The Novel ESR adds sedimentation and packing phase information obtained by hematology analyzer measurands. High correlation with WG was ensured by predicting the ESR value using Hematocrit (Hct) and MCV values as correcting parameters. METHODS: Novel ESR was compared with Modified WG (MONITOR-40, Joko Corporation) and Reference WG, according to internationally recognized guidelines: Precision, carryover, limit of quantification, comparability, linearity, accuracy, and fibrinogen sensitivity. Samples from healthy volunteers and clinical patients were used. The correction performance of Novel ESR and Modified WG was compared with Reference WG by regression analysis in three range categories for ESR and measurands affecting ESR correction (Hct, MCV, and MCH). RESULTS: Novel ESR showed sufficient basic performance and comparability with Modified WG. In the accuracy study comparing with Reference WG, the regression equation was y = 1.026x + 0.5(r =  .945,P <  .001;n = 271). When evaluating the correction performance, the slopes were within 0.8-1.2, except for the high part of Hct. All intercepts were within 10 mm. CONCLUSION: This study validated the correction performance to the initial estimated ESR value by aggregation phase information using information reflecting sedimentation and packing phase obtained from automated hematology analyzer. The Celltac α+ Novel ESR provided results equivalent to Reference WG.

The Current Status of Screening for Essential Thrombocythemia and Polycythemia Vera in Clinical Practice-Report from a Single Institution.

OBJECTIVE: We investigate the current status of screening for essential thrombocythemia(ET)and polycythemia vera(PV), at our hospital. METHODS: According to the World Health Organization(WHO)diagnostic criteria. PATIENTS: All patients who visited Juntendo University Urayasu Hospital between May 1984(when the hospital opened)and January 2019. RESULT: More than 90% of patients with elevated platelet counts(PLT)(n=25,062)and more than 90% of patients with elevated hemoglobin( Hb)or hematocrit(Ht)levels(n=16,422)did not visit the department of hematology, suggesting that there could be a high percentage of patients with potentially latent ET and PV visiting the hospital. In addition, a large number of patients fulfilling the laboratory criteria for ET/PV visited various departments of the hospital other than the department of hematology. CONCLUSION: Because ET/PV manifests with diverse symptoms, including non-specific symptoms and symptoms pertaining to other organ systems. Based on the findings, we consider that it is essential to disseminate information about the WHO diagnostic criteria/clinical symptoms and possibility of latent ET/PV to all departments of the hospital, and to establish cooperation between the department of hematology and other departments.

Threshold for optimal administration of plerixafor in autologous peripheral blood stem cell collections through CD34+ cell monitoring based on the experience from two Japanese university hospitals.

Plerixafor was introduced to Japan in 2017 as a stem cell mobilization enhancement reagent, but the threshold for its use remains unclear. In this study, we assessed 57 patients treated with plerixafor (33 patients with multiple myeloma (MM) and 24 with malignant lymphoma (ML) and 152 patients without plerixafor administration. When CD34+ cell pre-counts were between 5.5 and 20 cells/µL in MM or 6 and 21 cells/µL in ML, the CD34+ cell count increased significantly, attaining the highest yield in response to plerixafor (achievement rate by one leukapheresis is 93.3% and 91.7% in MM and ML, at P < .001 and P = .012, respectively). In case the CD34+ cell pre-count was less than 5.5 cells/µL, an increase of at least 7 cells/µL from baseline by plerixafor was the necessary condition to achieve successful collection through a two-time leukapheresis. Monitoring CD34+ cell numbers might improve the collection efficiency and reduce the cost.

Genomic analysis of antibiotic resistance for Acinetobacter baumannii in a critical care center.

AIM: Acinetobacter baumannii is commonly associated with outbreaks and antibiotic-resistant nosocomial infection. This study aimed to determine the relationship between antibiotic resistance and genotypes of A. baumannii. METHODS: A study was undertaken in the critical care center (CCC) of Juntendo University Urayasu Hospital (Urayasu, Japan) between January 2012 and September 2015. Antimicrobial susceptibility tests were carried out according to the Clinical and Laboratory Standards Institute guidelines. All A. baumannii isolates were verified to carry carbapenemase genes and the ISA ba1 element using polymerase chain reaction. The genetic relationship of all A. baumannii isolates was determined by pulsed-field gel electrophoresis and multilocus sequence typing. RESULTS: During the study period, 1634 patients were admitted to the CCC. Acinetobacter baumannii was detected in 43 patients (average age, 58 ± 19 years; 67.4% men). Six patients were determined to be extensively drug-resistant A. baumannii and 21 patients determined to be multidrug-resistant A. baumannii. Antimicrobial susceptibility linked genotypes of A. baumannii. Molecular characterization by pulsed-field gel electrophoresis and multilocus sequence typing showed that closely related clones of A. baumannii had spread in the CCC. CONCLUSION: Resistance to antimicrobial drugs was significantly associated with certain A. baumannii genotypic types and molecular types. Thus, we might be able to predict whether the genotype has spread in the CCC or not when the susceptibility is examined, facilitating the appropriate isolation of patients.

Marked thrombocytopenia in a neonate is associated with anti-HPA-5b, anti-HLA-A31, and anti-HLA-B55 antibodies.

Maternal antibodies against human platelet antigen (HPA) and/or human leukocyte antigen (HLA) cause fetal and neonatal alloimmune thrombocytopenia (FNAIT) in 0.09-0.15% of live births. Severe cases account for 5-31% and the frequency of multiple kinds of alloantibodies is 6.9-9% of FNAIT. We present a case of severe FNAIT associated with anti-HPA-5b, anti-HLA-A31, and anti-HLA-B55 antibodies, successfully treated with immunoglobulin and platelet transfusion. The anti-HLA-B55 antibody was detected in the newborn's serum, but disappeared on the 20th day, which was followed by an increase of the platelet count. These findings suggested the potential involvement of an anti-HLA antibody in the pathogenesis of FNAIT.

Pancreatic lipase activity in overnight effluent predicts high transport status in peritoneal dialysis patients.

BACKGROUND: Long-term peritoneal dialysis (PD) causes peritoneal morphological and functional changes, resulting in high transport status featuring increased peritoneal permeability. High transport status is diagnosed by peritoneal equilibration test (PET), a reliable but time-consuming method. We identifed a reliable biomarker in peritoneal effluent to predict high transport status in PD patients. METHODS: We collected peritoneal effluent and serum from 33 PD patients and measured common laboratory test parameters. High transport status was determined by PET if the dialysate/plasma ratio of creatinine at 4h dwell (D/P Cr 4h) was ≥0.81. RESULTS: There were significant correlations between D/P Cr 4h and some laboratory parameters in overnight effluent (pancreatic lipase activity, r=0.65, p<0.001; ß2-microglobulin concentration, r=0.59, p<0.001; IL-6 concentration, r=0.53, p<0.001; and CA125 concentration, r=0.29, p=0.027). In a multivariate logistic regression analysis, the pancreatic lipase activity in overnight effluent was identified as an independent predictor of high transport status even after adjusting for age, PD duration, and glomerular filtration rate [OR=1.43 (95% CI: 1.11-1.83), p=0.005]. CONCLUSIONS: The pancreatic lipase activity in overnight effluent is an independent predictor of high transport status in PD patients.

[Utilization of Multi-Institutional Laboratory Data as an Evidence Database: The Current Status and Future Tasks--Chairmen's Introductory Remarks].

The construction of a database that integrates raw laboratory data and diagnostic information with patient backgrounds is an effective tool in the practice of Evidence-Based Laboratory Medicine (EBLM). By exploring this type of database, it is possible to understand the diagnostic characteristics of the tests for a specific patient subgroup or condition. Although several studies have been carried out recently, these databases contain single-hospital data, and are thus limited regarding their external validity. In order to improve the reliability of the evidence, joint multi-institutional research is required. Therefore, the EBLM Committee of the Japanese Society of Clinical Laboratory Medicine arranged the symposium, entitled: "Utilization of multi-institutional laboratory data as an evidence database", which discusses current problems and solutions for the integration of multi-institutional laboratory data. In the symposium, five speakers presented on the following subjects: 1) Standardization of laboratory test coding (JLAC10); 2) The construction of a data warehouse in the hospital; 3) Multi-institutional study on long-term data changes; 4) Multi-institutional study on diagnostic accuracy; and 5) The construction of databases for the practice of EBLM and the need for the standardization/harmonization of laboratory data.

Mean postprandial triglyceride concentration is an independent risk factor for carotid atherosclerosis in patients with type 2 diabetes.

BACKGROUND: Postprandial hypertriglyceridemia is a risk factor for atherosclerotic disease. However, the postprandial triglyceride (PTG) concentration fluctuates markedly and is poorly reproducible. The aim of this study was to determine whether the mean PTG (mean-PTG) concentration is a risk factor for carotid atherosclerosis in patients with type 2 diabetes. METHODS: We measured the fasting and postprandial lipid concentrations, and the maximum intima-media thickness (max IMT) of carotid arteries by ultrasound in 115 diabetic patients. A carotid plaque was defined as max IMT of >1.0mm. The mean-PTG concentration was calculated from several PTG concentrations measured on different days during a 1-year follow-up period. RESULTS: PTG concentrations showed marked intra-individual variability, and ranged from 0.29 to 6.03 mmol/l. Patients with carotid plaques had higher mean-PTG concentrations than those without carotid plaques (1.51 ± 0.57 vs. 1.29 ± 0.47 mmol/l, p=0.025). Neither fasting triglycerides nor one-point PTG concentrations differed between the two groups. Multivariate stepwise logistic regression analysis revealed that the mean-PTG concentration was significantly associated with carotid plaques [OR 1.20 (95% CI, 1.05-1.37), p=0.009], even after adjusting for traditional risk factors including HDL-cholesterol, LDL-cholesterol, age, hypertension, and duration of diabetes. CONCLUSIONS: The mean-PTG concentration is an independent risk factor for carotid atherosclerosis in patients with type 2 diabetes.

Performance evaluation of the automated morphological analysis of erythrocytes by CellaVision DM96.

BACKGROUND: Automated digital morphology systems are utilized for blood cell morphological examination. The aim of this study is to evaluate the accuracy and efficacy of RBC morphological anomaly screening using the CellaVision DM96 (DM96) automated image analysis system. METHODS: The automated analysis of RBC shape, size, and chromasia abnormalities was conducted on the DM96 using 478 blood samples. A manual microscopic review was independently performed. RESULTS: The DM96 preclassified samples as poikilocytosis-positive for 98% of cases with schistocytosis or echinocytosis, 97% of elliptocytosis, and 92% or 65% of cases that were positive for teardrop cells or for target cells, respectively. The accuracy of the DM96 in the detection of RBC size and chromasia abnormalities of iron deficiency anemia cases was higher than direct microscopic observation. CONCLUSIONS: Automated morphological analysis with the DM96 has potential utility in the morphological screening of RBC anomalies that are associated with disease.

Association between antimicrobial consumption and clinical isolates of methicillin-resistant Staphylococcus aureus: a 14-year study.

The objective of this study was to determine the relationship between clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) and antimicrobial consumption in hospitalized patients over a 14-year period. The study was retrospectively conducted between January 1995 and December 2008 at Juntendo University Hospital, Tokyo, Japan, a 1,020-bed tertiary-care teaching hospital. The incidence of MRSA isolates was examined using clinical specimens presented to the microbiology laboratory in the hospital. Antimicrobial consumption through intravenous injection was calculated in terms of the number of defined daily doses per 100 bed-days. The correlation between the incidence of MRSA isolates and antimicrobial consumption was determined employing a multiple stepwise regression analysis. A total of 109,946 bacterial isolates were consecutively collected over the 14-year period, and, of these, 13,872 (64% of S. aureus strains excluding coagulase-negative staphylococci) were MRSA strains. The longitudinal observation showed that the number and rate of MRSA isolates marginally decreased. The rate of MRSA isolates among S. aureus strains in 1995 was 68.5%, whereas that in 2008 was 53.8%. Consumption of cephalosporins decreased. Among carbapenems, the rate of imipenem (IPM) consumption decreased, whereas that of meropenem increased. A multiple stepwise regression analysis revealed that the antimicrobial consumption of cefmetazole, cefotiam, and IPM was positively correlated with the incidence of MRSA isolates. The use of ß-lactam antimicrobials may contribute to the development of MRSA strains.

The use of CellaVision competency software for external quality assessment and continuing professional development.

AIMS: Quality assessment of blood cell morphological testing, such as white blood cell (WBC) differential and its interpretation, is one of the most important and difficult assignments in haematology laboratories. A monthly survey was performed to assess the possible role of the proficiency testing program produced by CellaVision competency software (CCS) in external quality assessment (EQA) of the clinical laboratories of affiliated university hospitals and the effective utilisation of this program in continuing professional development (CPD). METHODS: Four monthly proficiency surveys were conducted in collaboration with four clinical laboratories affiliated with the teaching hospitals of Juntendo University of Medicine in Japan. RESULTS: EQA results by the CCS proficiency testing program revealed a difference of performance levels of WBC differential and morphological interpretation and a discrepancy in the WBC differential criteria among laboratories. With regard to the utilisation of this proficiency program as a tool for CPD, this program successfully improved the performance of the low-scoring laboratories and less experienced individuals. CONCLUSIONS: The CCS proficiency testing program was useful for the quality assessment of laboratory performance, for education, and for the storage and distribution of cell images to be utilised for further standardisation and education.

Identification of Bcl-2/IgH fusion sequences using real-time PCR and chip-based microcapillary electrophoresis.

BACKGROUND: The determination of polymerase chain reaction (PCR) amplification product sizes of the Bcl-2/IgH fusion gene from follicular lymphoma (FL) provides evidence of clonal identity. METHODS: The present study describes detection of Bcl-2/IgH fusion gene clonality utilizing a small, simple microcapillary electrophoretic chip combined with a real-time PCR method. RESULTS: The microcapillary electrophoretic chip system effectively detects size differences among the Bcl-2/IgH fusion gene amplification products of FL from patient samples; something that is not possible using traditional gel electrophoresis. We also describe the potential of this system to utilize formalin-fixed, paraffin-embedded tissue samples sectioned on charged slides. CONCLUSIONS: The simple detection of Bcl-2/IgH fusion gene clonality using a microcapillary electrophoretic chip provides reliable information for monitoring minimal residual disease of FL, and can be an effective tool for use in clinical laboratories.

[Evaluation of capability of cell count and detection of tumor cells in cerebrospinal and body fluids by automated hematology analyzer].

Sysmex XE-5000 offers the body fluid modus which provides the opportunity to count and differentiate leukocytes in body fluids and cerebrospinal fluid (CFS). In this study, we evaluated the basic performance of this application using routinely obtained samples in comparison with manual counting. Reproducibility study yielded good results in samples with a high white blood cell (WBC) count, whereas relatively high imprecision was observed at low WBC counts. Linearity was established up to 1,500 cells/microL in CFS and 5,600 cells/microL in body fluid. The cell count by XE-5000 was highly correlated with that of the microscopic reference method. Highly fluorescent body fluid cells percent (HF-BF%) was observed in samples with tumor cells or activated macrophages, which provides information about the possible presence of tumor cells. In conclusion, total and differential WBC counts in body fluid and CFS can be reliably determined by XE-5000 in samples with increased cell counts. XE-5000 also provides screening information about the presence of tumor cells for further manual examination.

[Serum zinc concentration decreases with age and is associated with anemia in middle-aged and elderly people].

Zinc (Zn) is an essential trace element for humans and its deficiency can lead to several clinical problems. This study examined the relationship between the serum zinc concentration and anemia in middle-aged and elderly people. Samples were obtained from 150 men and 303 women who received health checkups over the course of 40 years. The serum concentration of Zn was measured as well as the complete blood count (CBC), alanine aminotransferase (AST), aspartate aminotransferase (ALT), gamma-glutamyl transferase (gamma GT), total cholesterol (TC), high density lipoprotein (HDL-C), triglyceride (TG), creatinine (Cr), uric acid (UA) and fasting plasma glucose (FPG). The serum Zn concentration was 77.4 +/- 9.7 microg/dL in men and 79.1 +/- 10.4 microg/dL in women (p= 0.09). The serum Zn concentration correlated inversely with age (r=-0.11, p=0.018). Anemia diagnosed by the World Health Organization criteria, was present in 17.3% of men and in 13.5% of women. However, more than 80% of the anemia was normocytic (men 86%, women 81%). The serum Zn concentration was significantly lower in those with anemia than in those without anemia. The Hb level correlated with the serum Zn concentration (men r=0.25, p=0.002, women r=0.23, p<0.001). A multiple regression analysis confirmed a low serum Zn concentration to be associated with a low Hb level. In conclusion, this study indicates that the serum Zn concentration decreases with age and that a low Zn concentration is associated with normocytic anemia, thus suggesting that a Zn deficiency may therefore be one of the causes of anemia in elderly people.

[Relevance of urinary protein/creatinine ratio and urinary abnormal casts].

OBJECTIVES: The quantification of 24 hrs urinary protein excretion is valuable for diagnosing and monitoring renal disease. However, because of its practical difficulties, the spot urinary protein/creatinine (P/C) ratio has been utilized. We aimed to evaluate the analytical performance of P/C ratio by comparing with the qualitative urinary protein values and the microscopic urine sediment analysis. METHODS: We obtained 5,538 urinary samples from the outpatients of Juntendo University Hospital. Testing for urinary P/C ratio was performed by Atlas Pro12 (cut-off 150 mg/g x Cr), urinary protein (proteinuria) was detected quantitatively by full-automated system ATLAS XL (cut-off 30 mg/dL). Microscopic exams were conducted following to the JCCLS reference method. RESULTS: The P/C ratio demonstrated higher sensitivity but lower specificity for urinary abnormal casts detected by microscopic exams compared to proteinuria (sensitivity; P/C 87%, proteinuria 77%. specificity; P/C 74%, proteinuria 93%). From the comparative study with microscopic exams, both P/C and proteinuria performed high positive rate (> 80%) for the granular cast type and mixture cast type. For the cellular cast type, however, the positive rate of P/C was 56% and that of proteinuria was only 36%. The overall abnormal casts by microscopic exams showed better correlation with the positive P/C ratio than proteinuria. CONCLUSION: This study emphasizes that a spot urine P/C ratio is useful in screening for the further microscopic exams. P/C ratio can be a convincing index of urinary protein excretion when attenuation urine is doubted.

High-sensitivity CRP reflects insulin resistance in smokers.

AIM: The elevation of high-sensitivity C-reactive protein (hs-CRP) is a strong risk factor for cardio-vascular disease (CVD) and is associated with insulin resistance. The hs-CRP concentration also increases in smokers known to be at a high risk for CVD. We examined whether hs-CRP concentra-tion reflects insulin resistance in smokers. METHODS: The hs-CRP levels were measured in 121 male subjects (54 nonsmokers and 67 smokers) with a normal glucose tolerance. The hs-CRP concentration was compared to the homeostasis model assessment of insulin resistance (HOMA-IR) and other clinical variables related to insulin resistance. RESULTS: Smokers had a 64.5% higher hs-CRP concentration than nonsmokers (p<0.0001). In both nonsmokers and smokers, hs-CRP positively correlated with HOMA-IR (r=0.301, p<0.05 and r=0.312, p<0.01) and fasting insulin (r=0.281, p<0.05 and r=0.356, p<0.01). The correlation between hs-CRP and HOMA-IR or fasting insulin was stronger in smokers than in nonsmokers. In smokers, hs-CRP significantly correlated with BMI and HDL-cholesterol (r=0.386, p<0.01 and r=-0.307, p<0.05). Stepwise regression analysis revealed that BMI and HOMA-IR were significant predictors of hs-CRP in smokers (r=0.423, p<0.01). CONCLUSIONS: The hs-CRP concentration reflects insulin resistance in smokers. It would be preferable to consider insulin resistance in evaluating hs-CRP concentrations, even in smokers.

Seasonal variation in liver function tests: a time-series analysis of outpatient data.

BACKGROUND: Long-term physiological variations, such as seasonal variations, affect the screening efficiency at medical checkups. This study examined the seasonal variation in liver function tests using recently described data-mining methods. METHODS: The 'latent reference values' of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), gamma-glutamyltransferase (gammaGT), cholinesterase (ChE) and total bilirubin (T-Bil) were extracted from a seven-year database of outpatients (aged 20-79 yr; comprising approximately 1,270,000 test results). After calculating the monthly means for each variable, the time-series data were separated into trend and seasonal components using a local regression model (Loess method). Then, a cosine function model (cosinor method) was applied to the seasonal component to determine the periodicity and fluctuation range. A two-year outpatient database (215,000 results) from another hospital was also analysed to confirm the reproducibility of these methods. RESULTS: The serum levels of test results tended to increase in the winter. The increase in AST and ALT was about 6% in men and women, and was greater than that in ChE, ALP (in men and women) and gammaGT (in men). In contrast, T-Bil increased by 3.6% (men) and 5.0% (women) in the summer. The total protein and albumin concentrations did not change significantly. AST and ALT showed similar seasonal variation in both institutions in the comparative analysis. CONCLUSIONS: The liver function tests were observed to show seasonal variations. These seasonal variations should therefore be taken into consideration when establishing either reference intervals or cut-off values, which are especially important regarding aminotransferases.