Development of a Novel Flow Cytometry-Based System for White Blood Cell Differential Counts: 10-color LeukoDiff.

BACKGROUND: Flow cytometry (FCM) is commonly used to identify many cell populations. We developed a white blood cell (WBC) differential counting system for detecting abnormal cells using FCM incorporating 10 colors and 11 antibodies in a single tube, called "10-color LeukoDiff," and evaluated its performance. METHODS: Ninety-one EDTA-anti-coagulated peripheral blood samples from 76 patients were analyzed using 10-color LeukoDiff. We compared 10 color LeukoDiff results with the results of manual differential count (manual diff). WBCs were classified into 17 cell populations: neutrophils, total lymphocytes, T lymphocytes, B lymphocytes, CD5 and CD19 co-expressing lymphocytes, natural killer cells, total monocytes, 16+ monocytes, eosinophils, immature granulocytes, basophils, myeloblasts, B-blasts, T-blasts, myeloid antigen-positive B-blasts, CD19- plasma cells, and 19+ plasma cells. RESULTS: The correlations between the 10-color LeukoDiff and manual diff results were strong (r>0.9) for mature neutrophils, lymphocytes, eosinophils, immature granulocytes, and blasts and moderate for monocytes and basophils (r=0.86 and 0.74, respectively). There was no discrepancy in blast detection between 10-color LeukoDiff and manual diff results. Furthermore, 10-color LeukoDiff could differentiate the lineage of the blasts and separately count chronic lymphocytic leukemic cells and multiple myeloma cells. CONCLUSIONS: The 10-color LeukoDiff provided an accurate and comprehensive WBC differential count. The most important ability of 10-color LeukoDiff is to detect blasts accurately. This system is clinically useful, especially for patients with hematologic diseases, such as acute leukemia, chronic lymphocytic leukemia, and multiple myeloma. Application of this system will improve the development of FCM gating strategy designs.

Clinical progress of human papillomavirus genotypes and their persistent infection in subjects with atypical squamous cells of undetermined significance cytology: Statistical and latent Dirichlet allocation analysis.

The present study aimed to investigate differences in prognosis based on human papillomavirus (HPV) infection, persistent infection and genotype variations for patients exhibiting atypical squamous cells of undetermined significance (ASCUS) in their initial Papanicolaou (PAP) test results. A latent Dirichlet allocation (LDA)-based tool was developed that may offer a facilitated means of communication to be employed during patient-doctor consultations. The present study assessed 491 patients (139 HPV-positive and 352 HPV-negative cases) with a PAP test result of ASCUS with a follow-up period ≥2 years. Patients underwent PAP and HPV DNA chip tests between January 2006 and January 2009. The HPV-positive subjects were followed up with at least 2 instances of PAP and HPV DNA chip tests. The most common genotypes observed were HPV-16 (25.9%, 36/139), HPV-52 (14.4%, 20/139), HPV-58 (13.7%, 19/139), HPV-56 (11.5%, 16/139), HPV-51 (9.4%, 13/139) and HPV-18 (8.6%, 12/139). A total of 33.3% (12/36) patients positive for HPV-16 had cervical intraepithelial neoplasia (CIN)2 or a worse result, which was significantly higher than the prevalence of CIN2 of 1.8% (8/455) in patients negative for HPV-16 (P<0.001), while no significant association was identified for other genotypes in terms of genotype and clinical progress. There was a significant association between clearance and good prognosis (P<0.001). Persistent infection was higher in patients aged ≥51 years (38.7%) than in those aged ≤50 years (20.4%; P=0.036). Progression from persistent infection to CIN2 or worse (19/34, 55.9%) was higher than clearance (0/105, 0.0%; P<0.001). In the LDA analysis, using symmetric Dirichlet priors α=0.1 and ß=0.01, and clusters (k)=5 or 10 provided the most meaningful groupings. Statistical and LDA analyses produced consistent results regarding the association between persistent infection of HPV-16, old age and long infection period with a clinical progression of CIN2 or worse. Therefore, LDA results may be presented as explanatory evidence during time-constrained patient-doctor consultations in order to deliver information regarding the patient's status.

Development of a cervical cancer progress prediction tool for human papillomavirus-positive Koreans: A support vector machine-based approach.

OBJECTIVES: To develop a Web-based tool to draw attention to patients positive for human papillomavirus (HPV) who have a high risk of progression to cervical cancer, in order to increase compliance with follow-up examinations and facilitate good doctor-patient communication. METHODS: Records were retrospectively analysed from women who were positive for HPV on initial testing (before any treatment). Information concerning age, Papanicolaou (PAP) smear result and presence of 15 high-risk HPV genotypes was used in a support vector machine (SVM) model, to identify the patient features that maximally contributed to progression to high-risk cervical lesions. RESULTS: Data from 731 subjects were analysed. The maximum number of correct cancer predictions was seen when four features (PAP, HPV16, HPV52 and HPV35) were used, giving an accuracy of 74.41%. A web-based high-risk cervical lesion prediction application tool was developed using the SVM model results. CONCLUSIONS: Use of the web-based prediction tool may help to increase patient compliance with physician advice, and may heighten awareness of the significance of regular follow-up HPV examinations for the prevention of cervical cancer, in Korean women predicted to have heightened risk of the disease.

The effect of thioctic acid on allodynia in a rat vincristine-induced neuropathy model.

OBJECTIVE: To investigate the antiallodynic effects of thioctic acid in vincristine-induced neuropathy in rats. METHODS: Neuropathy was induced in Sprague-Dawley rats via vincristine intraperitoneal injection. After 15 days, rats were investigated for the presence of mechanical and cold allodynia, and those with allodynia received intraperitoneal injection with normal saline or 1, 5, or 10 mg/kg thioctic acid. Mechanical and cold allodynia were assessed before treatment and at 15, 30, 60, 90, 150 and 180 min after treatment. RESULTS: Mechanical and cold allodynia were reduced by thioctic acid injection. The duration of effect increased with thioctic acid dose. CONCLUSION: Thioctic acid may be an effective treatment for vincristine-induced neuropathy.

Flow cytometric white blood cell differential using CytoDiff is excellent for counting blasts.

BACKGROUND: The usefulness of the CytoDiff flow cytometric system (Beckman Coulter, USA) has been studied in various conditions, but its performance including rapidity in detecting and counting blasts, the most significant abnormal cells in the peripheral blood, has not been well evaluated. The objective of this study was to evaluate the performance of the CytoDiff differential counting method in challenging samples with blasts. METHODS: In total, 815 blood samples were analyzed. Samples flagged as "blasts" or "variant lymphocytes" and showing <10% blasts by manual counts were included. In total, 322 samples showed blasts on manual counts, ranging from 0.5% to 99%. The CytoDiff method was performed by flow cytometry (FC500; Beckman Coulter, USA) with a pre-mixed CytoDiff reagent and analyzing software (CytoDiff CXP 2.0; Beckman Coulter). RESULTS: The average time required to analyze 20 samples was approximately 60 min for manual counts, and the hands-on time for the CytoDiff method was 15 min. The correlation between the CytoDiff and manual counts was good (r>0.8) for neutrophils and lymphocytes but poor (r<0.8) for other cells. When the cutoff value of the CytoDiff blast count was set at 1%, the sensitivity was 94.4% (95% CI; 91.2-96.6) and specificity was 91.9% (95% CI; 89.0-94.1). The positive predictive value was 88.4% (95% CI; 84.4-91.5) (304/344 cases) and negative predictive value was 96.2% (95% CI; 93.9-97.7) (453/471 cases). The CytoDiff blast counts correlated well to the manual counts (r=0.9223). CONCLUSIONS: The CytoDiff method is a specific, sensitive, and rapid method for counting blasts. A cutoff value of 1% of at least 1 type of blast is recommended for positive CytoDiff blast counts.

A novel marker for screening paroxysmal nocturnal hemoglobinuria using routine complete blood count and cell population data.

BACKGROUND: Final diagnosis of paroxysmal nocturnal hemoglobinuria (PNH) may take years demanding a quick diagnosis measure. We used the facts that PNH cells are damaged in acid, and reagents for measuring reticulocytes in Coulter DxH800 (Beckman Coulter, USA) are weakly acidic and hypotonic, to create a new PNH screening marker. METHODS: We analyzed 979 complete blood counts (CBC) data from 963 patients including 57 data from 44 PNH patients. Standard criteria for PNH assay for population selection were followed: flow cytometry for CD55 and CD59 on red blood cells (RBCs) to a detection level of 1%; and fluorescent aerolysin, CD24 and CD15 in granulocytes to 0.1%. Twenty-four PNH minor clone-positive samples (minor-PNH+) were taken, in which the clone population was <5% of RBCs and/or granulocytes. Excluding PNH and minor-PNH+ patients, the population was divided into anemia, malignancy, infection, and normal groups. Parameters exhibiting a distinct demarcation between PNH and non-PNH groups were identified, and each parameter cutoff value was sought that includes the maximum [minimum] number of PNH [non-PNH] patients. RESULTS: Cutoff values for 5 selected CBC parameters (MRV, RDWR, MSCV, MN-AL2-NRET, and IRF) were determined. Positive rates were: PNH (86.0%), minor-PNH+ (33.3%), others (5.0%), anemia (13.4%), malignancy (5.3%), infection (3.7%), normal (0.0%); within anemia group, aplastic anemia (40.0%), immune hemolytic anemia (11.1%), iron deficiency anemia (1.6%). Sensitivity (86.0%), specificity (95.0%), PPV (52.1%), and NPV (99.1%) were achieved in PNH screening. CONCLUSION: A new PNH screening marker is proposed with 95% specificity and 86% sensitivity. The flag identifies PNH patients, reducing time to final diagnosis by flow cytometry.

Novel markers of early neutrophilic and monocytic engraftment after hematopoietic stem cell transplantation.

BACKGROUND: Numerous studies tried to find new markers that after hematopoietic stem cell transplantation predict engraftment earlier than the conventional marker, absolute neutrophil count (ANC >500/µL). Early engraftment prediction can be achieved by a marker that reflects the release of neutrophils and monocytes into the leukopenic peripheral blood. METHODS: We analyzed blood cell parameters, including cell population data such as volume, conductivity, and light scatter in 77 patients who underwent HSCT (allogeneic, n=63; autologous, n=11) to detect possible markers. RESULTS: We identified 2 early engraftment markers of neutrophils (NEUTRO) and monocytes (MONO); a pair of mean-volume-neutrophils (MNV) and mean-conductivity-neutrophils (MNC) for NEUTRO; and a pair of mean-volume-monocytes (MMV) and mean-conductivity-monocytes (MMC) for MONO. The new markers showed distinct patterns for early engraftment wherein 1) on the engraftment day, MNV peaked as MNC notched simultaneously for every case, and 2) MMV peaked as MMC notched simultaneously in most cases. Engraftment was predicted 3.8±2.7 days earlier than by ANC in 74 successful engraftment cases by using NEUTRO and/or MONO: 1) 72 cases (97.3%), in which NEUTRO and/or MONO predicted earlier engraftment than ANC, 2) 1 case, in which the 3 markers predicted engraftment on the same day, and 3) 1 case, in which NEUTRO predicted engraftment on the same day as ANC and MONO failed to predict engraftment. CONCLUSIONS: By analyzing the data from daily complete blood counts, engraftment can be predicted approximately 4 days earlier than ANC >500/µL using NEUTRO as a base marker and MONO as a supplementary marker.

Clinical validation of AdvanSure GenoBlot assay as primary screening and test of cure for human papillomavirus infection.

BACKGROUND: Clinical specificity and sensitivity are essential factors in the adoption of a human papillomavirus (HPV) test as a primary screening tool and test of cure after treatment of cervical cancer and precancerous lesions (High-Risk-Lesion). Using histologically-confirmed High-Risk-Lesion-patient specimens with postoperative follow-ups, we performed clinical validation of the AdvanSure GenoBlot Assay (GenoBlot; LG Life Sciences, Korea). METHODS: The study population included 100 cases with High-Risk-Lesion, 96 with high-risk genotype positive and cervical intraepithelial neoplasia (CIN) 1 or better, and 39 with HR-negative and better than CIN 1. Forty-eight High-Risk-Lesion cases received follow-up HPV exams after surgery. For validation as a test of cure, 48 preoperative specimens (PreOP) and 78 postoperative specimens (PostOP) from 48 subjects were separately analyzed. The results of HPV DNA chip tests (HPVDNAChip; BioMedLab Co., Korea) and sequencing were cross-compared. RESULTS: The concordance rates for each genotype between HPVDNAChip and GenoBlot were between 96.3-100%. The accuracy of HPVDNAChip and GenoBlot was 87.9% and 96.6%, respectively. Genotype-based specificity for High-Risk-Lesion detection was higher than 87% for both assays; genotype 16 showed the highest sensitivity. In the PostOP group, the positive rates for HPVDNAChip and GenoBlot were 30.8% and 47.4%, respectively. CONCLUSIONS: GenoBlot showed a higher positive rate than HPVDNAChip for each genotype, with concordance rate and accuracy being similar to previous reports. As a test of cure, GenoBlot performed better than the HPVDNAChip.

Comparison of the AdvanSure HBV real-time PCR test with three other HBV DNA quantification assays.

BACKGROUND: We compared the AdvanSure hepatitis B virus real-time polymerase chain reaction (AdvanSure HBV) kit with three other HBV DNA quantification assays and evaluated its performance. METHODS: The AdvanSure HBV real-time PCR assay was compared with the Abbott RealTime HBV Quantification Kit, the COBAS TaqMan HBV Test, and the VERSANT HBV branched DNA 3.0 assay. The precision, linearity, accuracy, limit of detection (LOD), cross reactivity, and genotype inclusivity of the assays were compared, and any influence of the sampling tube type was evaluated. RESULTS: The AdvanSure HBV PCR showed good correlations with the three other HBV DNA assays. The R(2) coefficients were 0.944, 0.939, and 0.921 with the Abbott RealTime HBV Quantification Kit, the COBAS TaqMan HBV Test, and the VERSANT bDNA 3.0 assay, respectively. Linearity was good in the tested range of 1.15-8.45 log10 IU/ml. The lower LOD result was consistent with the 18 IU/ml claimed by the manufacturer. HBV genotypes A-F were all correctly amplified, and no cross reactivity was found in samples with high HCV RNA levels or high protein concentrations. The results were not influenced by the sample preparation tube (i.e. plain tube, SST, and EDTA containing tubes). CONCLUSION: The AdvanSure HBV real-time PCR assay is a reliable method for quantifying HBV DNA levels in routine laboratory testing.

Comparative study of immunofluorescent antinuclear antibody test and line immunoassay detecting 15 specific autoantibodies in patients with systemic rheumatic disease.

Based on the currently proposed algorithms, antibodies specificities (sp-ANAs) are identified mainly in samples positive for fluorescent antinuclear antibodies (FANA) screening tests. The purpose of the present study was to compare diagnostic performances of FANA and line immune assay (LIA) detecting 15 sp-ANAs in patients with systemic rheumatic diseases (SRD). In 948 sera from the patients with SRD (n = 590) and non-SRD (n = 358), we evaluated the fluorescent patterns and intensities in the FANA test, and compared the FANA results with sp-ANAs against nRNP, Sm, SS-A, Ro52, SS-B, Scl-70, PM/Scl, Jo-1, CENP B, PCNA, dsDNA, nucleosome, histone, ribosomal-P, and M2. The sensitivity and specificity was 75.9% and 52.5% of FANA test and 62.0% and 84.4% of sp-ANAs test for SRD detection. The overall agreement between FANA and sp-ANAs results was 69.2% (Kappa coefficient; 0.404). According to the clinical diagnosis, the levels of agreement varied from 33.3% to 83.1%. The positive predictive values of each FANA pattern for the detection of sp-ANAs were less than 50% except for the discrete speckled pattern (91.7%). The 1:100 intensity of FANA as well as the monoreactivity of LIA, anti-SSA(-)/anti-Ro52(+), or FANA(-)/sp-ANAs(+) was associated with non-SRD. Antibodies against ribosomal-P or PCNA were specific for systemic lupus eryhthematosus. This study highlights the need for careful interpretation of FANA test results to assess sp-ANAs and the application of sp-ANAs tests including less-common autoantibodies. In patients with clinical suspicion of SRD, screening with both FANA and sp-ANAs tests could improve diagnostic efficiency.

[The relationship between the timing of gestational diabetes screening and HbA1c level and neonatal outcome].

BACKGROUND: The aim of this study was to observe clinical outcomes of the mother and her infant who were possibly exposed to high blood glucose at least 2-3 months in the early and midterm pregnancy by checking gestational weeks (GW) and the first HbA1c level at initial diagnosis of gestational diabetes (GDM). METHODS: A total of 107 GDM patients and their newborns were subject of this study. GDM patients were newly diagnosed at the Holy Family Hospital of Catholic University from January 2003 until December 2007 and continuously managed in the diabetes center. Patients medical records were retrospectively reviewed to evaluate GW and HbA1c level at the time of diagnosis, and clinical outcomes of mother and newborn baby. RESULTS: The proportion of subjects who had been diagnosed of having GDM according to GW was 7.5%, in less than 24th week of pregnancy; 55.1% in the 24-28th week; 28.0% in the 29-32nd week; and 9.4% 33rd week or more. There were 39 out of 107 subjects (36.4%) with HbA1c levels >or=6.5% and 26 out of 39 subjects (24.3%) with HbA1c levels >or=7.0%. In clinical outcomes of newborn by HbA1c levels, the frequency of delivery of large for gestational age (LGA) infant was higher in mothers diagnosed with GDM after 29th week of pregnancy or with HbA1c levels 7.0% or more (P<0.001). CONCLUSIONS: If the screening test for gestational DM was delayed, HbA1c level and the risk for LGA seemed to be higher, so it may be necessary to screen GDM no later than 24th week of pregnancy.

Comparison of culture screening protocols for methicillin-resistant Staphylococcus aureus (MRSA) using a chromogenic agar (MRSA-Select).

To compare the culture screening protocols for methicillin-resistant Staphylococcus aureus (MRSA), a total of 300 duplicate nasal swabs (233 initial cultures and 67 weekly follow-up cultures) were collected consecutively from 233 patients in the Intensive Care Unit (ICU). One swab was plated directly on MRSA-Select agar (D-MRSA-Select) and observed at 24 hr. The duplicate swab was incubated in tryptic soy broth (TSB) with 6.5% NaCl for 24 hr, and then subcultured on MRSA-Select (B-MRSA-Select), BAP (B-BAP), and mannitol salt agar with 4 mg/L oxacillin (B-MSA(OXA)), and observed at 24 hr. MRSA was detected in 13.7% (32/233) of the initial and 22.4% (15/67) of the follow-up specimens. A patient was classified as MRSA-positive if any of the media grew colonies that were tested and confirmed to be MRSA. In the initial screening samples, the sensitivities of D-MRSA-Select, B-MRSA-Select, B-BAP, and B-MSA(OXA) were 78.1%, 84.4%, 78.1%, and 65.6%, respectively, and the specificities were 100%, 98.0%, 83.1%, and 93.5%, respectively. The sensitivities of all but the B-MRSA-Select protocol were significantly lower (p <0.05). In follow-up screening, the sensitivities of D-MRSA-Select, B-MRSA-Select, B-BAP, and B-MSA(OXA) were 66.7%, 86.7%, 66.7%, and 53.3%, respectively, and the specificities were 100%, 98.1%, 90.4%, and 90.4%, respectively. D-MRSA-Select protocol was considered useful in screening for MRSA because it was fast, highly specific, and showed sensitivity comparable to B-BAP. Salt-containing enrichment broth in conjunction with MRSA-Select (B-MRSA-Select) provides a promising way to increase sensitivity in initial and follow-up screening for MRSA.

[Distribution of antigenic aberration in the bone marrow of acute leukemia in complete remission].

BACKGROUND: The aberrant, leukemia-associated antigen expression patterns allow us to discriminate leukemic blasts from normal precursor cells. Our major goal was to determine a guideline for the detection of minimal residual disease using CD20+/CD34+ and myeloid Ag+/CD19+ combination in the bone marrow of acute leukemia in complete remission (CR) after chemotherapy. METHODS: Bone marrow samples from 117 patients with acute leukemia in complete remission after chemotherapy and from 22 healthy controls were immunophenotyped by triple staining and measured by flow cytometry. RESULTS: The CD20+/CD34+ cells in the large lymphocyte gate (R1) ranged from 0% to 3.% (0.8plusmn;0.82%, P=0.000) in CD20+/CD34+ B-lineage ALL CR (N=31), from 0.03% to 4.2% (0.7plusmn;0.83%, P=0.000) in CD20-/CD34- B-lineage ALL CR (N=66), from 0.1% to 0.96% (0.45plusmn;0.32%, P=0.016) in T-ALL CR (N=10), and from 0.02% to 0.48% (0.18plusmn;0.15%, P=0.776) in AML CR (N=10). The CD13,33+/CD19+ cells in R1 gate ranged from 0% to 2.69% (0.37plusmn;0.48%, P<0.001) in CD13,33+/CD19+ B-lineage ALL CR (N=31), from 0% to 1.8% (0.31plusmn;0.28%, P<0.001) in CD13,33-/CD19+B-lineage ALL CR (N=65), from 0.02% to 0.64% (0.29plusmn;0.22%, P=0.071) in T-ALL CR (N=9), and from 0% to 0.17% (0.07plusmn;0.09%, P=0.341) in AML CR (N=3). CONCLUSIONS: Using an immunophenotypic method for the detection of early relapse or minimal residual disease of B-lineage ALL bone marrow in CR after chemotherapy, different cutoff values should be applied according to antigen combination and gating. When the proportion of aberrant antigen combination was less than 5% in large lymphocyte gate, the results should be interpreted with caution.

[Clinical efficacy of HPV DNA chip test in the era of HPV vaccination: 1,211 cases, a single institution study].

BACKGROUND: Human papillomavirus (HPV) prophylactic vaccines, bivalent types for HPV-16/18 with 70% prophylactic expectation, have been developed based on the genotypes found prevalent in the western countries, but little is known for those in Korea. Using a DNA chip test, we evaluated the clinical efficacy of HPV genotype based on cervical abnormalities. METHODS: As the initial diagnostic tests, HPV DNA chip tests and Papanicolaou smear (PAP) were used for 1,211 subjects. Cervical colposcopy directed biopsies were performed for 626 among the 1,211 subjects within one month. RESULTS: The most frequently found genotypes in all HPV-positive specimens (n=445) were HPV-16 (22.0%), 58 (13.9%), 52 (11.0%), 51 (9.0%), 56 (8.5%), and 18 (7.2%). HPV prevalence was significantly higher in specimens where PAP and biopsy results were closer to malignancy. The HPV genotype distribution of the histologically confirmed cervical high-grade squamous intraepithelial lesions (HSIL) or carcinoma cases showed HPV-16, 58, 52, 18, and 33, in descending order. The HPV DNA chip sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for the detection of cervical HSIL or carcinoma were 76.9%, 70.1%, 72.1%, and 75.8%, respectively, Of these, the sensitivity and NPV were higher than those of PAP. PPV and NPV of HPV-16 were 90.5% and 60.7%, respectively, being the highest among the genotypes. CONCLUSIONS: We confirmed that HPV-16 genotype was also very important for the diagnosis of HSIL and cervical carcinoma in Korea. However, contrary to the findings in the western countries, the prevalence of HPV-58 was higher than that of HPV-18. Moreover, as the other HPV genotype reports were rare in Korea, further studies are required with the HPV DNA chip test before the nationwide adoption of the vaccines.

Quantitative comparisons of antibody-binding sites of platelet glycoprotein IIb/IIIa in aplastic anemia and idiopathic thrombocytopenic purpura.

The numbers of antibody-binding sites of platelet glycoprotein (GP) IIb/IIIa on circulating platelets were analyzed using 4 kinds of antibodies in 34 aplastic anemia (AA) patients, 20 idiopathic thrombocytopenic purpura (ITP) patients, and 14 normal controls. The numbers of antibody-binding sites of CD41, CD41a, CD41b, and CD61 on platelets of the AA patients were less than in the normal controls (p <0.001). In the ITP patients, the numbers of sites for CD41 and CD41a were less than in normal controls (p <0.05). There were significant positive correlations between CD41 and CD41a, CD41b, and CD61 in the 3 groups. There were significant negative correlations between CD41 and CD41b and between CD41a and CD41b in the normal controls, but not in the AA or ITP patients. In summary, the numbers of the 4 antibody-binding sites of GPIIb/IIIa on platelets of AA and ITP patients are different from those in normal controls. Measurements of the antibody-binding sites of GPIIb/IIIa are not necessary for the differential diagnosis of AA and ITP. However, the differences in correlations between the numbers of epitopes in AA and ITP patients suggest that the epitopes of GPIIb/IIIa are altered in these diseases.

[A case of IgA kappa light chain deposition disease and combined adult Fanconi syndrome with auer rod-like intracytoplasmic inclusions in plasma cells and proximal renal tubular cells].

We report a case of IgA kappa light chain deposition disease and combined adult Fanconi syndrome with Auer rod-like intracytoplasmic inclusions in plasma cells and proximal renal tubular cells in a 54-yr-old female. Cytochemical stainings revealed a strong acid phosphatase activity of the inclusions and weak periodic acid-Schiff positivity, whereas the reactions for peroxidase and alpha-naphthyl acetate esterase were negative. An immunostaining verified IgA-kappa inside the plasma cells. Kidney biopsy revealed Bence Jones cast nephropathy with kappa light chain positivity, and Congo red staining was negative. Electron microscopy showed needle-shaped crystals located in tubular epithelial cells.

[Proportions of cells expressing CD38-/CD34+, CD38+/CD34+, CD19+/CD34+, or CD13,33+/CD34+ in the regenerating bone marrows during complete remission of acute leukemia or after bone marrow transplantation].

BACKGROUND: The hemopoietic stem cells increase in number during the regeneration after chemotherapy or bone marrow transplantation (BMT). Although the proportion of hemopoietic stem cells and their differentiation have been studied by immunophenotyping using the flow cytometry, no substantial research efforts have been directed toward the regenerating marrow. We attempted to discover the proportions of undifferentiated stem cells, committed stem cells, B cell precursors, and myeloid precursors in the regenerating bone marrows during complete remission (CR) and after engraftment of BMT. METHODS: Bone marrow samples from 82 patients with acute leukemia in CR and from 25 patients after BMT engraftment, along with 22 control samples, were used to find the numbers of CD38-/CD34+, CD38+/CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells in the large lymphocyte gate by flow cytometry. We cross-analyzed our results in terms of groups: CR, BMT, and initial diagnosis groups. We performed significance tests on age, relapse, chromosomal abnormalities, clinical outcomes, and initial immunophenotypes of the leukemic cells. RESULTS: The proportions of CD38-/CD34+, CD38+/CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells are more highly distributed in acute B-lymphoblastic leukemia than the normal group and also in the CR than the BMT group. CD19+/CD34+ cells were increased in the relapse group and CD38+/ CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells were increased in the group with chromosomal abnormality. The results were irrelevant to the initial immunophenotype of the leukemic blasts. CONCLUSIONS: The increases of the markers spanned too widely to apply one specific cutoff value to analyze them. They seemed to be the results of normal regeneration, irrelevant to relapse or initial immunophenotype of leukemic blasts.

Diagnostic value of Elecsys S100 as a marker of acute brain injury in the emergency department.

The aim of this study was to evaluate the screening and diagnostic validity of the initial serum S100 levels by Elecsys S100 immunoassay in an emergency department. We measured serum S100 in 101 acute brain injury (ABI) patients, in 40 healthy subjects, and in 41 chronic brain injury (CBI) patients (the control group). The ABI patients were divided in several groups according to the Glasgow Coma Scale (GCS), injury type and findings of brain imaging. Of the 101 patients, the 66 cranial computed tomography (CCT) or brain magnetic resonance imaging (MRI)-positive patients showed higher S100 levels than did the 35 CCT/MRI-negative patients (P=0.028). At a cutoff of 0.105 microg/L, the sensitivity was 84.8% and the specificity 74.3% for all of the ABI patients, and the sensitivity was increased to 96.9% for the traumatic ABI patients for detecting image-positive ABI. There was no significant difference of S100 levels among the groups on the CCT or MRI pathological findings (P=0.478). In conclusion, serum S100 levels can be used in the emergency department as an additional screening tool to identify CCT- or MRI-positive ABI.

Comparison of protocols for surveillance of methicillin-resistant Staphylococcus aureus (MRSA): medical staff vs ICU patients.

To compare the sensitivity of various protocols for methicillin-resistant Staphylococcus aureus (MRSA) surveillance, active surveillance for detecting MRSA nasal colonization was performed on 97 members of the medical staff and 218 patients in the Intensive Care Unit (ICU) of a university hospital. Duplicate nasal swabs were collected from each participant. One was plated directly on a blood agar plate (D-BAP) and observed at 24 and 48 hr. Another was incubated overnight in tryptic soy broth (TSB) with 6.5% NaCl, and subcultured on both BAP (B-BAP) and mannitol salt agar with 4 mg/L of oxacillin (B-MSAOXA). The MRSA colonization rate was similar in the medical staff and patient samples (16.5% vs 11.9%, p = 0.285). Among the medical staff members, the sensitivity of MRSA detection was the same (93.8%) in D-BAP and B-BAP. In the ICU patients, which are a high-risk group, the sensitivity of MRSA detection was improved by adding a pre-enrichment step (73.1% on D-BAP vs 96.2% on B-BAP). The simple direct plating protocol was sufficiently sensitive for the medical staff members, but pre-enrichment was an essential step to increase detection of MRSA in the ICU patients.