BACKGROUND: Chlamydia pneumoniae is a recently recognized species consisting of the strains commonly referred to as TWAR. These strains are associated with acute respiratory infections in humans, especially atypical pneumonia. So we tried to make a monoclonal antibody to Chlamydia pneumoniae. METHODS: C. pneumoniae were adapted to grow in HeLa-229 cells. The organisms were harvested and purified in a linear gradient of renograffin. BALB/c mice (female, 10weeks) were intravenously immunized with purified C. pneumoniae(TW-183).The spleen cells and SP 2/0 myeloma cells were fused with 40% polyethylene glycol (Mol.Wt.:1,450). Antibodies against C. pneumoniae were screened by an enzyme- linked immunosorbent assay (ELISA). The proteins of purified chlamydial elementary bodies were separated by discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and Western blots were performed with these monoclonal antibodies. RESULTS: Two monoclonal antibodies (HYMD10, HYMG12) reacted specifically with C. pneumoniae, as measured by an ELISA and indirect immunofluorecent stain. One of the monoclonal antibody (HYMD10) reacted with 75- and 39-KDa proteins in Western blot. The other monoclonal antibody (HYMG12) reacted with 98- and 39-KDa proteins of C. pneumoniae. CONCLUSIONS: These species-specific monoclonal antibodies (HYMD10, HYMG12) to C. pneumoniae could be used for diagnosis of C. pneumoniae infections.
Animals ; Antibodies ; Antibodies, Monoclonal ; Blotting, Western ; Chlamydia* ; Chlamydophila pneumoniae* ; Diagnosis ; Diatrizoate Meglumine ; Electrophoresis ; Enzyme-Linked Immunosorbent Assay ; Humans ; Mice ; Pneumonia ; Polyethylene Glycols ; Respiratory Tract Infections ; Sodium ; Spleen

Inositol-1,4,5-triphosphate [IP3] receptors binding protein released with IP3 (IRBIT) was previously reported as an activator of NBCe1-B. Recent studies have characterized IRBIT homologue S-Adenosylhomocysteine hydrolase-like 2 (AHCYL2). AHCYL2 is highly homologous to IRBIT (88%) and heteromerizes with IRBIT. The two important domains in the N-terminus of AHCYL2 are a PEST domain and a coiled-coil domain which are highly comparable to those in IRBIT. Therefore, in this study, we tried to identify the role of those domains in mouse AHCYL2 (Ahcyl2), and we succeeded in identifying PEST domain of Ahcyl2 as a regulation region for NBCe1-B activity. Site directed mutagenesis and coimmunoprecipitation assay showed that NBCe1-B binds to the N-terminal Ahcyl2-PEST domain, and its binding is determined by the phosphorylation of 4 critical serine residues (Ser151, Ser154, Ser157, and Ser160) in Ahcyl2 PEST domain. Also we revealed that 4 critical serine residues in Ahcyl2 PEST domain are indispensable for the activation of NBCe1-B using measurement of intracellular pH experiment. Thus, these results suggested that the NBCe1-B is interacted with 4 critical serine residues in Ahcyl2 PEST domain, which play an important role in intracellular pH regulation through NBCe1-B.
Animals ; Carrier Proteins ; Hydrogen-Ion Concentration ; Mice ; Mutagenesis, Site-Directed ; Phosphorylation ; S-Adenosylhomocysteine ; Serine*

We report a case of Strongyloides stercoralis infection in a 85-year-old male patient who had complained of poor oral intake, diarrhea, and upper abdominal pain for 6 months. At admission, he showed severe eosinophilia in peripheral blood. Rhabditiform larvae were detected in the stool examination on the 15th admission day and developed into filariform larvae with a notched tail after stool culture by the Harada-Mori method. The patient received albendazole therapy for 7 days but no improvement were observed and he fell into pulmonary edema and coma.
Abdominal Pain ; Aged, 80 and over ; Albendazole ; Coma ; Diarrhea ; Eosinophilia* ; Humans ; Larva ; Male ; Pulmonary Edema ; Strongyloides stercoralis* ; Strongyloides*

BACKGROUND: The main causes of thrombocytopenia in cirrhosis are thought to be platelet destruction and the reduction of thrombopoietin (TPO) expression in the liver. Immature platelet fraction (IPF) has been measured by a fully automated analyzer (Sysmex XE-2100, Japan) as reticulated platelet (RP), which is reflected with thrombopoiesis in bone marrow. In this study, we tried to compare the percentage of IPF (IPF) with that of RP (RP) in patients with liver cirrhosis (LC) and controls. METHODS: We compared IPF to RP in 72 liver cirrhosis patients and 30 healthy normal controls. RP was stained with acridine orange, followed by FC500 (Beckman Coulter, USA) analysis and the IPF was identified by flow cytometry with the use of a nucleic acid specific dye in the reticulocyte channel on the Sysmex XE-2100 (TOA Medical Electronics Co., Ltd., Japan). RESULTS: IPF value in the healthy control was 2.2% (1.7-5.2). RP and IPF were significantly higher in the patients with liver cirrhosis (P<0.05). IPF appeared to be correlated with RP (y=0.19x+3.35, r=0.34, P<0.05). In ROC for diagnosis of LC, IPF was significantly more useful than RP. CONCLUSIONS: This results show that a rapid, inexpensive automated method for measuring the IPF is feasible and should become a standard parameter in evaluating reticulated platelets.
Aged ; Female ; Humans ; Liver Cirrhosis/*complications ; Male ; Middle Aged ; Platelet Count/*methods ; Stem Cells/*cytology ; Thrombocytopenia/*diagnosis/etiology

BACKGROUND: D-dimer is a specific marker of secondary fibrinolysis. D-dimer assay is widely used in the diagnosis of disseminated intravascular coagulation, deep vein thrombosis, pulmonary embolism, and arterial thromboembolism. Enzyme linked immunosorbent assays have been validated as the reference method for plasma D-dimer measurement, but it took long time. We evaluated the analytical performance of new automated coagulation system, ACL TOP, for quantification of D-dimer. METHODS: The total plasma D-dimer concentrations were measured by Nycocard and ACL TOP. To test the linearity, a serial dilution samples were prepared and measured. Between run precision of the ACL TOP D-dimer assay was evaluated with HemosIL D-Dimer controls for 20days. The correlation was evaluated using 75 plasma samples from patients. ACL TOP was evaluated according to CLSI guidelines. RESULTS: ACL TOP showed good linearity (r=0.9996) and between run coefficient of variation was within 4.0%. Coefficient of correlation between Nycocard and ACL TOP was 0.798. Positive concordance rate of ACL TOP was 67%, and negative concordance rate of ACL TOP was 80%. CONCLUSIONS: Since the ACL TOP showed a satisfactory precision, linearity, and comparative high correlation with Nycocard, and is more convenient and automatic than the Nycocard, it should be potentially beneficial in the clinical laboratories.
Diagnosis ; Disseminated Intravascular Coagulation ; Fibrinolysis ; Humans ; Plasma ; Pulmonary Embolism ; Thromboembolism ; Venous Thrombosis

Persistent bone marrow aplasia after intensive chemotherapy is uncommon, but is one of the fatal complications in patients with acute myeloid leukemia (AML). Although allogeneic hematopoietic stem cell transplantation (HSCT) is considered to be contraindicated for patients who have hematologic diseases with serious infections, such as bacterial septicemia or invasive fungal diseases, combined with prolonged neutropenia due to frequent morbidity and mortality, such risks can be overcome by non-myeloablative conditioning and best supportive care. Here, we report an AML patient with persistent marrow aplasia after induction therapy, treated successfully with reduced-intensity allogeneic HSCT despite severe bacterial and fungal infections.
Anemia, Aplastic ; Bone Marrow* ; Drug Therapy* ; Hematologic Diseases ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myeloid, Acute* ; Mortality ; Neutropenia ; Sepsis ; Stem Cell Transplantation* ; Stem Cells*

Thrombotic thrombocytopenic purpura (TTP) is a clinical syndrome characterized by micro-angiopathic hemolytic anemia, thrombocytopenia, fever, renal disorders, and neurological manifestations. Its clinical course is rapid and the mortality rate is high if untreated or relapse occurs. Previous studies reported that rituximab, a monoclonal antibody for CD20 surface antigen on B lymphocytes, may be effective in treating idiopathic TTP that is refractory to plasma exchange or relapses after remission. A 27-year-old Vietnamese man presented with fever and fatigue starting 3 days earlier, which was diagnosed as idiopathic TTP. To overcome his poor responsiveness to combined therapy using steroids and plasma exchange, rituximab was considered. In the current case, the patient was treated with a lower dose of rituximab, instead of the conventional 375 mg/m2/week, and achieved successful remission.
Adult ; Anemia, Hemolytic ; Antigens, Surface ; Asian Continental Ancestry Group ; B-Lymphocytes ; Fatigue ; Fever ; Glucocorticoids ; Humans ; Mortality ; Neurologic Manifestations ; Plasma Exchange ; Plasma* ; Purpura, Thrombotic Thrombocytopenic* ; Recurrence ; Steroids ; Thrombocytopenia ; Thrombotic Microangiopathies ; Rituximab

BACKGROUND: Inducible MLS(B) (macrolide-lincosamide-streptogramin B) resistance in staphylococci is not detected by standard susceptibility test methods. Failure to identify inducible MLS(B) resistance may lead to clinical failure during clindamycin therapy. We determined the prevalence of inducible MLS(B) resistance in erythromycin-resistant staphylococcal isolates. MATERIALS AND METHODS: We evaluated all 2,792 non-duplicate staphylococcal strains: 1,402 Staphylococcus aureus and 1,390 coagulase-negative staphylococci (CoNS) isolated from May 2008-June 2009 at one-unoversity hospital. Testing for inducible MLS(B) was accomplished by the disk approximation test (D-test) in accordance with the recommendations of the Clinical and Laboratory Standards Institute (CLSI). RESULTS: Of the 2,792 staphylococcal isolates, 892 S. aureus isolates and 740 CoNS isolates were resistant to erythromycin. Among the 892 erythromycin-resistant S. aureus isolates, the overall prevalence of inducible MLS(B) was 21.3% (16.2% of MRSA and 76.3% of methicillin-susceptible S. aureus). Among the 740 erythromycin-resistant CoNS isolates, the overall prevalence of inducible MLS(B) was 16.5% (16.0% of methicillin-resistant CoNS and 18.7% of methicillin-susceptible CoNS). The D-test was positive in 88.8% of S. aureus and 28.4% of CoNS isolates, which were erythromycin-resistant and clindamycin-susceptible. CONCLUSIONS: There are some variations in the prevalence of inducible MLS(B) resistance in clinical staphylococcal isolates. It is important that clinical laboratories report inducible MLS(B) resistance for erythromycin-resistant and clindamycinsusceptible staphylococcal isolates.
Clindamycin ; Erythromycin ; Methicillin Resistance ; Methicillin-Resistant Staphylococcus aureus ; Prevalence ; Staphylococcus ; Staphylococcus aureus

Background: The Sysmex DI-60 system (Sysmex, Japan) is an automated cell image analyzer. This study aimed to evaluate the performance of the DI-60 system for the differential analysis of leukocytes. Methods: A total of 220 samples were analyzed in this study. The agreement between DI-60 pre-classification and manual verification by experts was determined. The correlation between the differential leukocyte counts obtained using the DI-60 system and those manually obtained in the peripheral blood smears were determined. Results: The pre-classification agreement of DI-60 was 91.0%. The correlation coefficients of normal five-part differentials were 0.9163 (segmented neutrophils), 0.9017 (lymphocytes), 0.8533 (monocytes), 0.8345 (eosinophils), and 0.3505 (basophils). The sensitivity, specificity, positive predictive value, negative predictive value, and the efficiency of counting the abnormal cells, including blasts, promyelocytes, myelocytes, metamyelocytes, lymphocyte variants, and erythroblasts, were determined. The efficiency of the DI-60 system in counting the blasts, promyelocytes, myelocytes, metamyelocytes, lymphocyte variants, and erythroblasts was 99.5%, 100.0%, 95.9%, 96.5%, 98.6%, 100.0%, and 95.9%, respectively. Conclusions The pre-classification agreement of DI-60 was higher than that of previous studies. The correlation between the differential leukocyte counts obtained with the DI-60 system and those of manual counting was acceptable. The performance of DI-60 as a screening tool in clinical laboratories may be good; however, it is yet to replace manual slide review.

BACKGROUND: In the severe community-acquired pneumonia, it has been known that the immune status is occasionally suppressed. This study was performed to identify the immunologic markers related with the prognostic factors in severe community-acquired pneumonia. METHODS: 23 patients with severe community-acquired pneumonia were involved in this study, and divided into survivor (16) and nonsurvivor (7) groups. In this study, the medical history, laboratory tests(complete blood counts, routine chemistry profile, immunoglobulins, complements, lymphocyte subsets, cytokines, sputum and blood culture, urine analysis), and chest radiographs were scrutinized. RESULTS: 1) Both groups had lymphopenia(total lymphocyte count 995.6±505.7/mm2 in the survivor and 624.0±287.6/mm2 in the nonsurvivor group). 2) The T-lymphocyte count of the nonsurvivor group(295.9±203.0/mm2) was lower than the survivor group(723.6±406.5/mm2) (p<0.05). 3) The total serum protein(albumin) was 6.0±1.0(2.7±0.7) g/dl in the survivor and 5.2±1.5(2.3±0.8)g/dl in the nonsurvivor group. The BUN of the noncurvivor group(41.7±30.0mg/dl) was higher than that of the survivor group(18.9±9.8mg/dl)(p<0.05). The creatinine concentration was higher in the nonsurvivor group(1.8±1.0mg/dl) than that in the survivor group(1.0±0.3mg/dl)(p<0.05). 4) The immunoglobulin G level was higher in the survivor group (1433.0±729.5mg/dl) than in the nonsurvivor group(849.1±373.1mg/dl)(p<0.05). 5) The complement C3 level was 108.0±37.9mg/dl in the survivor group and 88.0±32.1mg/dl in the nonsurvivor group. 6) A cytokine study showed an insignificant differenne in both groups. 7) Chronic liver disease, DM, and COPD were major underlying diseases in both groups. CONCLUSION: These results suggest that decreased a T-lymphocyte count and immunoglobulin G level, and an increased BUN and creatinine level may be associated with the poor prognosis of severe community-acquired pneumonia.
Biomarkers ; Chemistry ; Complement C3 ; Complement System Proteins ; Creatinine ; Cytokines ; Humans ; Immunoglobulin G ; Immunoglobulins ; Immunologic Factors* ; Liver Diseases ; Lymphocyte Count ; Lymphocyte Subsets ; Pneumonia* ; Prognosis* ; Prospective Studies* ; Pulmonary Disease, Chronic Obstructive ; Radiography, Thoracic ; Sputum ; Survivors ; T-Lymphocytes