Haemolytic transfusion reactions due to Mi-antibodies; need to include MiltenbergerIII positive cells in pre-transfusion antibody screening in Hong Kong.
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Biomedical subjects
Publications and source records attributed to C K Lin.
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Low density lipoprotein receptors (LDL-R) on Daudi Burkitt's lymphoma (BL) cells were assessed using fluorescent DiI (3,3'-dioctadecylindocarbocyanine iodide)-LDL and flow cytometric analyses. Receptor-specific binding of DiI-LDL is followed by internalization of the bound complex and lysosomal hydrolysis of the ligand. Increase in the fluorescence intensity per cell is hence used as a measure of DiI-LDL uptake and, implicitly, as an indication of LDL-R presence. Our results show that uptake was observed in > 98% of the Daudi cells, and the level of uptake was significant and clearly distinguishable from autofluorescence, suggesting that: (a) this assay is comparable to the iodinated LDL uptake assay, although the ED50 values for the ligands are different; (b) this assay is comparable to the flow-cytometric detection of LDL-R using a commercial antibody directed against the receptor itself, and superior to a similar assay based on an antibody directed against membrane-bound LDL; (c) LDL uptake could be monitored along with transferrin uptake, suggesting that multiple endocytic receptor activities can be concurrently studied; (d) DiI-LDL uptake can be examined along with fluorescein-conjugated anti-CD10, -CD19, and -CD71, with little cross-interference, offering the added advantage that endocytic uptake and phenotyping can be simultaneously monitored; (e) the expression of LDL-R is intrinsically elevated in diverse cell lines such as Daudi, Raji, Ramos, Jurkat, and WIL2-NS, but not in normal lymphocytes. Our results therefore indicate that flow cytometric analysis of DiI-LDL uptake has potentially useful applications in the detection and study of endocytic receptor LDL-R in B and T lymphocytic cell lines.
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BACKGROUND: Sequential cycles of combination chemotherapy with high-dose cyclophosphamide, etoposide and cisplatin (sHDCEP) can largely increase the total dose (TD) of drug delivered. If granulocyte colony-stimulating factor (G-CSF) and/or autologous peripheral blood stem cell (PBSC) rescue can shorten the duration of cytopenia between cycles of sHDCEP, the dose intensity (DI) can be increased as well. In order to explore the feasibility of delivering maximal TD and DI by administration of sHDCEP with G-CSF and/or PBSC rescue, this trial is undertaken to investigate the hematologic and nonhematologic toxicity observed with sHDCEP by G-CSF and/or PBSC rescue. METHODS: Patients with refractory malignancy and well preserved physiologic function for whom no available therapy is likely to cure or prolong the survival were eligible for the study. Each cycle of high-dose chemotherapy consisted of: cyclophosphamide 5,000 mg/m2, etoposide 1,500 mg/m2 and cisplatin 150 mg/m2. G-CSF and/or PBSC were administered alternatively after each cycle as rescue for myelosuppression. The next cycle was given to patient who showed response to the previous cycle after recovery from toxicity for a maximal of 4 cycles. RESULTS: Two cases of refractory malignancy with progressive disease were treated by sHDCEP for 7 cycles, including 4 cycles with G-CSF rescue, 2 cycle with PBSC rescue, and 1 cycle with G-CSF + PBSC rescue. In the 4 cycles rescued by G-CSF alone, we observed a slightly slower granulocyte and markedly prolonged platelet recovery in the subsequent cycle. By comparing the effect of G-CSF and/or PBSC rescue on hematologic recovery with the preceding cycle in the same patient, we found that G-CSF rescue provided faster granulocyte recovery than PBSC, but PBSC rescue provided faster platelet recovery than G-CSF. Rescue by larger number of PBSCs provided only faster platelet but not granulocyte recovery than rescue by adding G-CSF to a very small number of PBSCs. However, G-CSF plus the very small number of PBSCs provided shorter duration of both granulocytopenia and thrombocytopenia than rescue by G-CSF alone. The most common nonhematologic toxicity from sHDCEP included transient nausea, vomiting, diarrhea and mild impairment of liver function but we observed no significant or irreversible major organ damage. The side effect from PBSC collection was mild and toxicity from reinfusion of the thawed PBSCs was not obvious. Using G-CSF and/or PBSC rescue, sHDCEP was delivered repeatedly in no more than 4 weeks for the next-cycles except for patient 1 who had cycle 4 delayed because of prolonged platelet recovery by only G-CSF rescue in cycle 3. CONCLUSIONS: Our initial experience has shown that the nonhematologic toxicity from sHDCEP, G-CSF and PBSC rescue was well tolerated. Prolonged platelet recovery after sequential cycles of HDCEP by only G-CSF rescue delayed the next cycle of chemotherapy. Although the next cycle was delivered within 4 weeks by only PBSC rescue, concurrent infusion of G-CSF and larger number of PBSCs should provide the most rapid hematologic recovery. Sequential high-dose chemotherapy administered by this model is likely to provide the maximal delivery of TD and DI, and is worthy of further clinical trials.
BACKGROUND: Patients with liver cirrhosis usually die of hepatic failure and variceal bleeding. Successful treatment of the latter can reduce mortality. Sclerotherapy is one method often used. This study compared (a) the successful rate of acute bleeding control; (b) short- and long-term survival rate between those with and without treatment with sclerotherapy to evaluate the clinical benefit of sclerotherapy for liver cirrhosis patients with esophageal variceal bleeding. METHODS: Between August 1983 and December 1991, 183 cirrhotic patients with esophageal variceal bleeding receiving endoscopic injection sclerotherapy (EIS) was compared with 123 patients without sclerotherapy treatment retrospectively. The severity of underlying liver disease was classified using a modified Child's classification. Sclerotherapy was done within 48 hours after active bleeding in the sclerotherapy-treated group, while the medical treatment group received Sengstaken-Blakemore (SB) tube or pitressin infusion only. RESULTS: Successful rate of acute bleeding control was 81.63% (120/147) in the EIS group and 59.35% (73/123) in the medical treatment group. The worse the hepatic function of the patients, the lower the success of acute bleeding control in both groups. Fifty subjects (74.63%) had varices eradicated in 67 sclerotherapy treatment patients with regular follow-up. Patients receiving EIS had a better long-term survival than those without treatment. Benefit of EIS on long-term survival was more significant in Child B patients and less in Child C and Child A patients. Death from variceal bleeding was lower in the EIS group than in the medical treatment group (32% vs 62.6%). Complications of EIS were rare. Eight patients died of aspiration pneumonia, spontaneous bacterial peritonitis or acute renal failure after sclerotherapy, and most were Child B and C patients. Sixteen patients had esophageal stricture. Four needed dilatation treatment. CONCLUSIONS: The sclerotherapy-treated group had a higher control rate of acute bleeding and lower mortality rate from esophageal variceal bleeding compared with the medical-treated group. The procedure prolonged long-term survival in Child B patients but did so less frequently in Child A and Child C patients. The incidence of complications was low. As a whole, EIS is a safe and efficient method for control of esophageal variceal bleeding.
Successful termination of a pregnancy and delivery of a normal infant in a young woman with Krukenberg tumor is reported. This association has rarely been described previously. The persistent gastrointestinal symptoms mimicking the early nausea and vomiting of pregnancy mask the presentation of a tumor in the stomach. Growth of the fetus leading to abdominal distension masks the presence of the metastatic ovarian tumor in the pelvic cavity. Thus, early diagnosis of the tumor may be delayed. We emphasize the importance of differentiation between physiological morning sickness and unusual persistent gastrointestinal symptoms in pregnant women. Persistent unusual gastrointestinal symptoms need careful evaluation by panendoscopic examination.
BACKGROUND: Lower concentration of ascitic or serum complement (C3, C4) or protein has been reported to participate in the development of spontaneous bacterial peritonitis (SBP). In Taiwan, the etiology of hepatic cirrhosis is mainly post-hepatic and SBP is the common complication. This study aims to determine the role of protein and complements in the pathogenesis of SBP. METHODS: 119 cirrhotic patients were divided into two groups, 30 SBP and 89 non-SBP. The concentrations of ascitic and serum complement and protein were measured for comparison. RESULTS: The ascitic and serum C3, C4 and protein levels were significantly lower (P < 0.05) in patients with SBP than in non-SBP patients. No significant differences were noted in the ascites/serum ratio of C3, C4 and protein in patient with or without SBP. CONCLUSIONS: Low levels of ascitic and serum protein and complements, C3 and C4, may be prone to develop SBP in our patients mostly with post-hepatitic cirrhosis.
BACKGROUND: Reticulocyte count plays a major role in anemic evaluation. The conventional method done by the manual supravital staining cannot subclassify the group of less than 2% of corrected reticulocyte count. The newly-developed flow cytometer provides different maturation fractions by measuring its fluorescent intensity. The reticulocytes are believed to shift to the circulation from the bone marrow earlier in more severe anemia. Therefore, the purpose of this study is to evaluate the role of reticulocyte maturation fractions and bone marrow reticulocyte in anemia classification. METHODS: By using a fully automated counter, the roles of the reticulocyte with maturation and their shifting from bone marrow were evaluated in anemias. Different groups of subjects (243 in total) including aplastic, nutritional, and infiltrative anemias and anemia due to excess destruction and blood loss were studied. Each subject had bone marrow examination for morphologic diagnosis and reticulocyte evaluation. RESULTS: Both the absolute count and the maturation fractions of reticulocytes showed significant difference among marrow infiltration, aplastic anemia, and hemolytic anemia. Both the absolute reticulocyte count and less mature fractions were lowest in aplastic group. The marrow reticulocyte counts and shift ratio to circulating blood added little benefit in the classification of anemias. CONCLUSIONS: The automated reticulocyte count with maturation fractions helps classify anemias, particularly for those with low reticulocyte count by the manual method.
Glycol ethers (including glycol ether esters) are a group of solvents with medium-high boiling points and low evaporation rates, possessing solvent characteristics of alcohol/ether functions (or ether/ester functions). They have been widely used in coatings and other industrial products for more than half a century. Recently, E-series glycol ethers have been found to show reproductive and teratogenic toxicity, and throughout much of the world they are being replaced by the so-called P-series glycol ethers. In responding to the impact of the worldwide transition from E- to P-series glycol ethers, the current status of glycol ether use in Taiwan was studied. This study focuses on the type and quantity of these solvents being used, worker and public knowledge about their hazards, and possible changes in government regulations being considered. In this study, we found that large quantities of E-series glycol ethers were imported and used in Taiwan. The best estimates are: 2-ME, 2,500 tons; 2-EE, 1,200 tons; 2-EEA, 5,000-8,000 tons; 2-BE, 8,000 tons annually in 1991. For P-series glycol ethers, only about 2,500 tons are being used. Lack of knowledge about the potential toxic effects of the E-series glycol ethers is very common among users, regulatory agencies, academic institutes, and the general public. It is hoped that the results of this study, along with educational efforts, government regulations, and provision of technical services, will help prevent Taiwan from becoming a dumping site for these toxic chemicals.
Using a fully automated reticulocyte counter, the roles of the reticulocyte count with maturation in pancytopenia were evaluated. Different groups of pancytopenia including aplastic anemia, infiltrative marrow disorder, hypersplenism, and megaloblastic anemia were recruited. All patients had bone marrow examinations for morphological diagnosis and reticulocyte evaluation using an automated counter. The roles of these parameters were then analyzed statistically in the differential evaluation among these conditions. The following subjects were studied: 292 normal subjects, 67 cases of aplastic anemia, 69 cases of marrow infiltration by different malignancies, 35 cases of hypersplenism, and 13 cases of megaloblastic anemia. The results showed that the absolute reticulocyte counts were lowest in the groups of aplastic anemia and megaloblastic anemia and highest in hypersplenism. Both showed significant differences from the infiltrative groups. The maturation fractions were most immature in the group of marrow infiltration and are significantly different from the other groups. It was concluded that the highest absolute reticulocyte count (> 0.09 10(12)/L) obtained in pancytopenic patients suggests it to be a case of hypersplenism. The lowest counts (< 0.03 10(12)/L) with lowest immature fractions (< 10%) suggest the groups of aplastic or megaloblastic anemias. The highest immature fractions (> 30%) with a nearly normal reticulocyte count favor the group of marrow infiltration.
We have explored the forms of the G protein-related beta subunit which are present in Daudi lymphoblastoid cells. Northern blotting with labeled beta-1 and beta-2 probes indicates that two messages of 3.3 kb and 1.7 kb are present for both beta-1 and beta-2, implying that multiple forms of the beta subunit are present. Antibodies were raised against two peptides of the beta subunit (residues 1-23 and 127-145). Both antibodies detected subunits at 35 kDa and 31 kDa, of which the 35-kDa form predominates in the membrane fraction and the 31-kDa one in the cell cytosol. Crosslinking of the membrane fraction with the cleavable crosslinker (DTSSP) caused a simultaneous diminution in the 31-kDa form while increasing the amount of the 35-kDa form--a pattern which was reversed upon the reduction of these crosslinks with DTT. Studies of the soluble form indicate that this is truly a soluble protein since centrifugation at 200,000 x g for 2 h did not diminish the levels of the protein in the soluble fraction. Sedimentation analysis indicates that the soluble beta-homologue is found in fractions which overlap with those which contain the mu chain of immunoglobulin at a position clearly distinct from the expected positions of free mu or free beta. Our results suggest that at least two forms of a subunit which is closely related to, or identical with, the beta subunit of G proteins are present in Daudi cells.
We have analysed the pattern of nucleotide sequence variability in the 5' non-coding region (5' NCR) of geographically dispersed variants of hepatitis C virus (HCV). Phylogenetic analysis of sequences in this region indicated the existence of a new virus type, provisionally termed type 4, the identity of which was confirmed by further analysis of the more variable part of the HCV core protein coding region. The geographical distribution of HCV type 4 was distinct from that of other HCV types, it being particularly widespread in Africa and absent or rare in Europe and the Far East. Much of the variability in the 5' NCR appears to be constrained by a requirement for specific secondary structures in the viral RNA. In one of the most variable regions of the 5' NCR (positions -169 to -114), most of the nucleotide changes that are characteristic of different HCV types were covariant, with complementary substitutions at other positions. According to the proposed secondary structure of the 5' NCR, such changes preserved base pairing within a stem-loop structure, whereas the nucleotide insertions found in a proportion of 5' NCR sequences, including those of type 4, localized exclusively to the non-base-paired terminal loop. The specific nucleotide substitutions in the 5' NCR that differentiate each of the four HCV types can be detected by restriction enzyme cleavage, providing a rapid and reliable method for virus typing.
The frequencies of HLA antigens were compared between 55 patients with and 38 patients without lymphocytotoxic antibodies formation after long-term platelet transfusions. Only HLA-B60 and B75 were found to manifest significant difference between these two groups. Patients with HLA homozygosity had a higher incidence of alloimmunization. Although most of the platelet alloantibodies were against HLA antigens of high frequency, the HLA-antibodies were induced at a rate different from the frequency of their corresponding antigens. The antibodies against the first and second HLA loci are of similar frequencies. In conclusion, the patients with HLA homozygous alleles have a higher incidence of platelet alloimmunization, and the antibody of certain specificities has higher rate of occurrence. These findings may be helpful in platelet-donor selection.
Pseudohypoparathyroidism type Ia (PHP-Ia), an inherited multi-hormone resistance syndrome, is associated with deficient cellular activity of the alpha-subunit of the guanine nucleotide-binding protein (Gs alpha) that stimulates adenylyl cyclase. We determined prevalence of three recently described mutations in exons 1 and 10 of the Gs alpha gene among 24 unrelated patients with PHP-Ia. Restriction analysis was used to detect two mutations that produce unique RFLPs, and allele-specific oligonucleotide hybridization was used to detect the other mutation. As none of these mutations were not found, genomic DNA was analyzed with denaturing gradient gel electrophoresis to screen for other mutations in exon 10. Mutations of the initiation codon and exon 10 in the Gs alpha gene thus rarely (< or = 4% each) cause PHP-Ia and the Gs alpha gene mutations causing PHP-Ia are heterogeneous and unique to each pedigree.
Lymphocytotoxicity test (LCT) and platelet suspension immunofluorescence test (PSIFT) were used together to screen platelet-associated antibodies in patients who received long-term platelet transfusion. Twenty-four of 53 patients (45.3%) were immunized subsequently. Since the concordance of LCT and PSIFT was 100%, most of the platelet associated antibodies were of HLA specificity, and platelet specific antibody alone (in absence of HLA) was not detected. The identified antibodies were anti-A2, A11, A24, B5, B46, B57, B60, and B62. The majority of them were against the high frequency HLA antigens in the Chinese population. The development of antibody could not be correlated with the number of platelet-donors exposed, the time interval after the initiation of platelet transfusion, or the percentage of reactive lymphocytotoxic panels. HLA antibody was the major factor in causing platelet alloimmunization in the Chinese patients. However, some other unknown factors should be looked for. In addition, ABO incompatibility did not affect the posttransfusional increment while the platelet was compatible with LCT crossmatching.
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We have explored the requirements for the induction of the N-formyl-methionyl-leucyl-phenylalanine (FMLP) response in Daudi cells after anti-immunoglobulin treatment. Our results indicate that (a) induction of responsiveness to FMLP was observed in Daudi only after crosslinking of surface immunoglobulin by anti-immunoglobulin; (b) this induced responsiveness was not observed in Ramos or Wil-2 cells; (c) the F(ab')2 fragment was sufficient for the induction of the FMLP response, but the Fab fragment and the Fc fragment were ineffective; (d) of the many agents active in B lymphocyte regulation which were tested, none were as effective as anti-immunoglobulin in the induction of the FMLP response; and (e) three inhibitors of calcium mobilization (W-7 (N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide), PMA (phorbol 12-myristate 13-acetate), and colchicine), acting on distinct mechanisms, inhibited both the calcium mobilization due to anti-immunoglobulin and the induction of responsiveness to FMLP. Our results suggest important determinants in the induction of a calcium-mobilizing FMLP response in cells of B lymphocyte lineage include (a) the cell type, (b) a selective requirement for activation via surface immunoglobulin, and (c) crosslinking of the surface immunoglobulin.
The reference range of serum alanine amino transferase (ALT) for the local population was established by testing 5,000 random voluntary Chinese blood donors of various age groups of both sexes. In addition, 1,769 serum samples with elevated ALT levels were also collected for anti-HCV assays using both the Abbott and Ortho anti-hepatitis C virus (HCV) assay kits. The relationship between serum ALT and anti-HCV tests was studied and the performances of both kits used were compared. It was found that while the prevalence of serum anti-HCV was 0.4% among hepatitis B surface antigen-negative donors with normal ALT, subjects with ALT between 2 and 3 standard deviations (SD) and greater than 3 SD above the mean level had respective prevalence of anti-HCV 3 and 9.5 times that of the normal ALT subjects. Both anti-HCV kits were found to identify in majority the same positive population among the different groups of subjects studied. In addition, it was observed that for subjects who were anti-HCV-positive, the higher the serum ALT level, the higher the mean anti-HCV ELISA ratio and this observation was similar for both anti-HCV kits used. We conclude that: (1) there is a direct relationship between serum ALT level and anti-HCV positivity by EIA; (2) there is a direct correlation between serum ALT level and anti-HCV ELISA ratio, and (3) both Abbott and Ortho anti-HCV kits perform similarly in the identification of positive serum samples.