OBJECTIVE: To investigate the procedure of pre-transfusion testing and transfusion strategy of patients with multiple myeloma (MM) treated by daratumumab (DarA). METHODS: The blood samples of MM patients before and after DarA treatment from the Fifth Affiliated Hospital of Sun Yat-sen University were collected, and the ABO/Rh blood group antigen identification and DAT test results were compared. The results of antibody screening and cross matching of the patients before and after inactivation of red blood cells with 0.2 mol/L dithiothreitol (DTT) were compared and analyzed. RESULTS: ABO/Rh blood group antigen typing showed no affecting in patients after treated by DarA; the result of DAT test showed negative. Irregular antibody screening showed that all the three cells(Ⅰ, Ⅱ and Ⅲ) were positive(1+~2+) and the self-control was negative. By microcolumn agglutination method, the main side of the multi-bag of blood showed no matched, while the secondary side showed all identical. After treated by DTT solution, the cross matching results in reagent red blood cells and the red blood cells of blood donors were both consistent, and the irregular antibody screening was negative. The K(+)O type erythrocytes used in parallel control were transformed into K(-)O type erythrocytes after DTT treatment. However, there was no significant changes in E(+) O type erythrocytes before and after DTT treatment. There was no condensation on the primary and secondary side of the condensed amine method. The primary and secondary sides of blood matching by saline method showed negative. CONCLUSION After treated by DarA, cross matching results from microcolumn agglutination method can be interfered by the residual drug antibody in MM patients, while the interference was eliminated in the presence of 0.2 mol/L DTT solution. However, no disturbance was observed when using condensed amine method or saline method. Therefore, corresponding transfusion procedures should be selected according to the emergency degree of blood transfusion to ensure the safety and timeliness of blood transfusion.
Antibodies, Monoclonal ; Blood Transfusion ; Dithiothreitol ; Humans ; Multiple Myeloma/therapy*

The fibrillar coat of Candida albicans is of interest as its significance in antigenicity, antiphagocytosis, and adherence to host tissues. The partial biochemical properties and ultrastructure of fibrillar coat induced by rabbit sera were examined. The induced fibrillar layer was destroyed by treatments of lyticase, proteinase K and dithiothreitol. The total protein concentration of fibrillar cell wall lysate was higher than that of non-fibrillar cell wall lysate, but the total sugar concentration was similar. On SDS-PAGE analysis, the protein profiles between in fibrillar cells and in non-fibrillar cells were shown to be different. In fibrillar cells, the major bands of cell wall lysate were 83, 66, 54, 47, 33, and 26 kDa in dithiothreitol-treated lysate. The proteins of 26 and 19 kDa were predominant in lyticase-treated lysate. Although the fibrillar thickness and protein amount of cell wall lysate were increased in according to the incubation time, the protein profiles did not changed. These results suggest that the proteins of 83, 66, 54, 47, 33, 26, and 19 kDa may be major constituents of fibrillar coat in C. albicans.
Candida albicans* ; Candida* ; Cell Wall ; Dithiothreitol ; Electrophoresis, Polyacrylamide Gel ; Endopeptidase K

BACKGROUND: Eosinophils play an important role in asthmatic airway inflammation collaborateing with other inflammatory cells. Nitric oxide (NO) may amplify and perpetuate allergic inflammation. OBJECTIVE: The present study was aimed to determine whether NO metabolites and eosinophil activation markers in sputum reflect the severity of asthma. METHODS: Sputum was obtained in 27 asthmatic patients. We processed freshly expectorated sputum separated from saliva and by a treatment with equal volume of dithiothreitol 0.1%, cytospins for cell count and special stain, we collected the supernatant for biochemical assay. We used immunocytochemical staining to detect EG2+ eosinophils, and fluoroimmunoassay to detect eosinophil cationic protein (ECP). NO metabolites were assayed by using modified Griess reaction. RESULTS: Moderate and severe asthmatic patients had a significantly higher proportion of eosinophils (37.9+/-7.8% vs 58.4+/-7.9% vs 8.9+/-2.0%, p<0.01) and lower proportion of macrophages (46.5+/-5.5% vs 18.1+/-4.6% vs 75.6+/-4.0%, p<0.01) compared to mild asthmatics. The proportion of EG2+ eosinophils, and ECP levels in the sputum were significantly higher in moderate and severe asthmatic patients than in mild asthmatic patients (p<0.01, respectively). The level of NO metabolites in the sputum was significantly increased in severe asthmatic patients compared to that in mild asthmatic patients (1372.0+/-168.5micromole/L vs 658.3+/-186.4micromole/L, p<0.05). The proportion of eosinophils in sputum was inversely correlated with FEV1 and FEV1/FVC. EG2+ eosinophils and ECP also had an inverse relationship with FEV1 and FEV1/FVC. CONCLUSION: These findings demonstrate that the proportion of eosinophils, ECP, EG2+ eosinophils, and NO metabolites in the sputum of patients with asthma may be correlated with asthma severity.
Asthma* ; Cell Count ; Dithiothreitol ; Eosinophil Cationic Protein* ; Eosinophils* ; Fluoroimmunoassay ; Humans ; Inflammation ; Macrophages ; Nitric Oxide* ; Saliva ; Sputum*

BACKGROUND AND OBJECTIVES: Inflammatory cells in nasal secretions could reflect pathologic conditions occurring in the mucosa of the nasal and paranasal sinuses. But the cytologic investigation of the nasal secretions has not been fully accepted as a clinical examination. The aim of this study was to evaluate the quantitive cytologic study of various nasal diseases and the clinical value of cytologic examination of nasal secretions. MATERIALS AND METHODS: We studied 8 patients with chronic sinusitis, 8 patients with nasal polyposis, 23 patients with allergic rhinitis, and 11 patients with hypertrophic rhinitis. The nasal secretions were treated with dithiothreitol and examined at a magnification of 400X under a light microscope. The differential cell count and frequency of appearance of individual cell type was evaluated. RESULTS: The majority of inflammatory cells were eosinophil in allergic rhinitis and hypertrophic rhinitis. In chronic sinusitis and nasal polyposis, the majority were neutrophil. Eosinophils and lymphocytes appear very frequently in nasal polyposis. CONCLUSION: The result suggests that the cytologic study of nasal secretion is a simple and useful method to determine cell populations under various conditions.
Cell Count ; Dithiothreitol ; Eosinophils ; Humans ; Lymphocytes ; Mucous Membrane ; Neutrophils ; Nose Diseases ; Paranasal Sinuses ; Rhinitis* ; Sinusitis*

BACKGROUND AND OBJECTIVES: Postnasal drip is one of the most common symptoms of chronic rhinosinusitis (CRS), and is the main cause of chronic cough. To evaluate the effect of upper airway inflammation defined as CRS on lower bronchial airway, we compared the cytology of nasal secretion (NS) and bronchoalveolar lavage fluid (BALF) of normal controls and patients with chronic rhinosinusitis accompanying with and without chronic cough normal controls. MATERIALS AND METHOD: Ten patients of CRS with postnasal drip were selected. Five of them had chronic cough and the others not. Five normal controls were selected. NS collected using Juhn's tymanic tap and BALF collected through fiberoptic bronchoscopy were diluted with dithiothreitol and PBS. These samples were centrifused and then cytospin slide was prepared. The cytology of the slides were evaluated under light microscope after Wright stain. To examine neutrophil activity, nitroblue tetrazolium dye (NBT) test was performd. Statistical analysis was performed using Mann-Whitney Rank Sum W test. RESULTS: In NS, there were no significant differences in the cell populations among coughing, noncoughing, and the normal control group. NBT positivity of coughing (34.2%) and noncoughing (31.5%) groups showed significantly higher than those of controls (8.6%). In BALF of coughing group, the population of macrophages (78.0%) was significantly lower than noncoughing (86.6%) and control (92.8%) groups, and population of lymphocytes (20.8%) was significantly higher than noncoughing (12.6%) and the control (6.4%) groups. In BALF of noncoughing group, the population macrophages was lower and those of lymphocytes were higher than the control group. CONCLUSION: These results suggest that CRS enhances increased local immune responses and decreased phagocytic activity of the lower airway. And chronic cough in patients with CRS is thought to be dependent on individual tolerance to cough provocation, not on aspiration of postnasal drip of discharge.
Bronchoalveolar Lavage Fluid* ; Bronchoalveolar Lavage* ; Bronchoscopy ; Cough* ; Dithiothreitol ; Humans ; Inflammation ; Lymphocytes ; Macrophages ; Neutrophils ; Nitroblue Tetrazolium ; Sinusitis

TREK (TWIK-RElated K+ channels) and TRAAK (TWIK-Related Arachidonic acid Activated K+ channels) were expressed in COS-7 cells, and the channel activities were recorded from inside-out membrane patches using holding potential of -40 mV in symmetrical 150 mM K+ solution. Intracellular application of an oxidizing agent, 5,5'-dithio-bis (2-nitrobenzoic acid) (DTNB), markedly decreased the activity of the TREK2, and the activity was partially reversed by the reducing agent, dithiothreitol (DTT). In order to examine the possibility that the target sites for the oxidizing agents might be located in the C-terminus of TREK2, two chimeras were constructed: TREK2 (1-383)/TASK3C and TREK2 (1-353)/TASK3C. The channel activity in the TREK2 (1-383)/TASK3C chimera was still inhibited by DTNB, but not in the TREK2 (1-353)/TASK3C chimera. These results indicate that TREK2 is inhibited by oxidation, and that the target site for oxidation is located between the amino acid residues 353 and 383 in the C-terminus of the TREK2 protein.
Animals ; Arachidonic Acid ; Chimera ; COS Cells ; Dithionitrobenzoic Acid ; Dithiothreitol ; Membranes ; Oxidants

The Luminex-based single antigen bead (SAB) assay is widely used to detect HLA antibody in transplant recipients. However, one limitation of the SAB assay is the prozone effect, which occurs mostly as a result of complement interference. We investigated the efficacy of EDTA treatment for overcoming the prozone effect and predicting C1q binding of HLA antibody. We subjected 27 non-treated (naïve) and EDTA-treated serum samples from highly sensitized patients to IgG-SAB assays, and we confirmed the prozone effect in 53% and 31% of class I and class II antibody tests, respectively, after EDTA treatment. When we conducted additional assays after dithiothreitol treatment and serum dilution, EDTA was the most efficacious in eliminating the prozone effect. Reducing the prozone effect by EDTA treatment strengthened the correlation between IgG mean fluorescence intensity (MFI) and C1q MFI values (ρ=0.825) as compared with the naïve sera (ρ=0.068). Although C1q positivity was dependent on the concentration of HLA antibody in EDTA-treated sera, the correlations varied individually. Overall, our results confirmed the efficacy of EDTA treatment for overcoming the prozone effect. EDTA treatment showed a positive effect on the correlation between IgG MFI and C1q MFI values.
Complement System Proteins ; Dithiothreitol ; Edetic Acid ; Fluorescence ; Humans ; Immunoglobulin G ; Transplant Recipients

OBJECTIVE: To investigate the effectiveness and safety of low concentration dithiothreitol (DTT) in removing the interference of monoclonal anti-CD38 on transfusion compatibility testing, and develop a reasonable clinical transfusion strategy. METHODS: The blood type, direct antiglobulin testing (DAT) and antibody screening were tested according to standard methods. Antibody screening cells and donor's red blood cells were treated by DTT 0.2, 0.1, 0.05, 0.02, 0.01 and 0.005 mol/L, and antibody screening and cross-matching of serums after monoclonal anti-CD38 treatment were performed by anti-human globulin card. RESULTS: The 0.01 mol/L DTT at 37℃ for 30 minutes could remove the effect of monoclonal anti-CD38 on antibody screening and cross-matching, meanwhile retain their effectiveness in detecting anti-K, anti-LW, anti-JMH, anti-Lu^b, anti-e, anti-Di^a and anti-Jk^a alloantibodies. All the 10 patients had no acute or delayed haemolytic transfusion reactions and their routine blood tests showed that the red blood cells transfusion was effective. CONCLUSION The 0.01 mol/L DTT is a safe and effective method for removing the interference of monoclonal anti-CD38 with transfusion compatibility testing, while retaining the ability to detect most alloantibodies.
Antibodies, Monoclonal/pharmacology* ; Blood Grouping and Crossmatching ; Blood Transfusion ; Dithiothreitol/pharmacology* ; Erythrocytes ; Humans ; Isoantibodies/pharmacology*

Several approaches have been introduced to detect allo-antibodies in the presence of warm auto-antibodies, and these methods include warm autoadsorption, cysteine-activated papain and dithiothreitol (ZZAP), and polyethylene glycol (PEG) and dilution of the patient's serum. Among them, the dilution technique is a simple and rapid method. During pretransfusion testing of a 33 year-old systemic lupus erythematosus (SLE) patient with warm auto-antibodies, antibody identification was done by the dilution technique with using serum diluted 1-in-8. The patient demonstrated an anti-Fy(b) pattern of reactivity in his sera. Contrary to our expectations, the phenotype of the erythrocytes was Fy(a+/b+) and the genotype, as assessed by performing PCR-restriction fragment length polymorphism (RFLP), was FY*A/FY*B. These results suggest that the antibody is an autoantibody showing anti-Fy(b) specificities. An antibody identification test using undiluted serum showed the same result when 40 days had passed. We report here on a case with auto-anti-Fy(b) proven by the dilution method in the presence of warm autoantibodies.
Autoantibodies ; Dithiothreitol ; Erythrocytes ; Genotype ; Humans ; Indicator Dilution Techniques ; Lupus Erythematosus, Systemic ; Papain ; Phenotype ; Polyethylene Glycols ; Sensitivity and Specificity*

LW antigens are expressed in higher intensities in D-positive blood cells than D-negative cells, which can result in false identification of anti-D in pretransfusion testing. Although several cases of anti-LW have been reported abroad, to the best of our knowledge, none have been reported in Korea. Herein, we report a case of anti-LW in a 58 year-old RhD positive patient with non-Hodgkin's lymphoma with a positive direct Coombs test and a suspicion of the presence of passive anti-D antibodies because of a history of intravenous immunoglobulin administration. However, during a 5-month follow up, the antibody was confirmed as anti-LW on grounds that it showed weakened reaction in dithiothreitol treated cells and enforced reaction in cord O+ cells when compared to the results from antibody identification panel cells.
Antibodies ; Blood Cells ; Coombs Test ; Dithiothreitol ; Follow-Up Studies ; Humans ; Immunoglobulins ; Korea ; Lymphoma, Non-Hodgkin ; Sensitivity and Specificity*