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Biomedical subjects

M Chan

Publications and source records attributed to M Chan.

At least 19 recordsLinked to original sources
PubMed1996

Natural Killer (NK) cell deficiency associated with an epitope-deficient Fc receptor type IIIA (CD16-II).

Susceptibility to herpes virus infections has been described in experimental animals depleted of NK cells and in patients with defective NK cell function. We have identified a child with recurrent infections, especially with herpes simplex virus, who had a decreased number of CD56(+)CD3(-) NK cells in circulation. Her NK cells expressed an altered form of the Fc receptor for IgG type IIIA (Fc gamma RIIIA or CD16-II) which was not reactive with the anti-CD16-II MoAb B73.1. Sequence analysis revealed the patient to be homozygous for a T to A substitution at position 230 of CD16-II cDNA, predicting a Leu(66) to His(66) change in the first immunoglobulin domain of CD16-II at the B73.1 recognition site. Spontaneous NK cell activity of the patient's peripheral blood mononuclear cells (PBMC) was markedly decreased, while antibody-dependent cellular cytotoxicity (ADCC) was unaffected. These results suggest that this child suffers from a defect affecting the development and function of NK cells, resulting in NK cytopenia and clinically significant immunodeficiency. The role of the CD16-II mutant in the pathogenesis of the patient's NK cell deficiency is discussed.

Antibodies, Monoclonal↗
PubMed1995

Expression of GPI-anchored complement regulatory proteins CD55 and CD59 differentiates two subpopulations of human CD56+ CD3- lymphocytes (NK cells).

CD55 and CD59 are both glycosylphosphatidylinositol (GPI)-anchored complement regulatory proteins found on the surface of most hemopoietic cells. Using three-color cytofluorographic analysis with antibodies recognizing CD56, CD3, and CD59 or CD55 we determined that CD56+CD3- lymphocytes (NK cells) expressed both CD55 and CD59 but at lower levels than CD3+ lymphocytes. Since the CD56+CD3- lymphocyte population is heterogeneous, we examined expression of CD55 and CD59 on selected CD56+CD3- lymphocyte populations by depleting peripheral blood leukocytes of T cells, B cells, and monocytes. Dual staining of the selected CD56+CD3- cells, which were > 95% CD56+, permitted the distiction of two subpopulations: a major CD16brightCD56dimCD55dimCD59dim subpopulation and a minor CD16dim/negCD56brightCD55brightCD59bright subpopulation. Treatment with phosphatidylinositol-specific phospholipase C released both the CD55 and CD59 antigens from the surface of CD56+CD3- cells, indicating that both are GPI-anchored, as they are on other lymphocytes. CD56+CD3- cell subpopulations were individually isolated by anti-CD55 or anti-CD16 negative selection and were functionally compared to the parent CD56+CD3- cell population. The CD16brightCD56dimCD55dimCD59dim cells killed NK targets well but did not proliferate well in response to rIL-2, whereas CD16dim/negCD56brightCD55brightCD59bright cells proliferated well in response to rIL-2 but did not kill NK targets efficiently. We conclude that all CD56+CD3- cells express some levels of the GPI-anchored proteins, CD55 and CD59, and that two CD56+CD3- subpopulations with different functional characteristics can be distinguished by the level of expression of these two antigens.

CD3 Complex↗
PubMed1995

Enhancement of phospholipid hydrolysis in vasopressin-stimulated BHK-21 and H9c2 cells.

The hydrolysis of phospholipids in vasopressin-stimulated baby hamster kidney (BHK)-21 and H9c2 myoblastic cells was investigated. Phosphatidylcholine and phosphatidylethanolamine in these cells were pulse labelled with [3H]glycerol, [3H]myristate, [3H]choline or [3H]ethanolamine, and chased with the non-labelled precursor until linear turnover rates were obtained. When cells labelled with [3H]glycerol or [3H]myristate were stimulated by vasopressin, no significant decrease in the labelling of phosphatidylcholine was detected, but the labelling of phosphatidic acid was elevated. However, the labellings of phosphatidylethanolamine and its hydrolytic product were not affected by vasopressin stimulation. When the cells were pulse labelled with [3H]-choline, vasopressin stimulation caused a decrease in the labelled phosphatidylcholine with a corresponding increase in the labelled choline. The apparent discrepancy between the two types of labelling might be explained by the recycling of labelled phosphatidic acid back into phosphatidylcholine, thus masking the reduction in the labelled phospholipid during vasopressin stimulation. Alternatively, the labelled choline produced by vasopressin stimulation was released into the medium, thus reducing the recycling of label precursor back into the phospholipid and making the decrease in the labelling of phosphatidylcholine readily detectable. Further studies revealed that vasopressin treatment caused an enhancement of phospholipase D activity in these cells. The presence of substrate-specific phospholipase D isoforms in mammalian tissues led us to postulate that the differential stimulation of phospholipid hydrolysis by vasopressin was caused by the enhancement of a phosphatidylcholine-specific phospholipase D in both BHK-21 and the H9c2 cells.

Animals↗
PubMed1995

Preliminary cryocrystallographic study of the mitochondrial cytochrome bc1 complex: improved crystallization and flash-cooling of a large membrane protein.

Ubiquinol-cytochrome c reductase is a crucial integral membrane protein in the mitochondrial respiratory cycle. Eleven subunits containing three cytochrome heme groups and a 2Fe-2S Rieske center make up this 240 kDa enzyme complex. Previously, many different crystal forms of the bc1 complex have displayed diffraction to as far as 4.5 A. However, rapid degradation of the protein in the X-ray beam at room temperature has obstructed the collection of a full data set from a single crystal. As slight heterogeneities between crystals severely hampered merging of data from different crystals, we sought a method to stabilize the protein crystal in the X-ray beam in order to collect a full data set from one crystal sample. To this end, water soluble protein crystals are frequently flash-cooled to cryogenic temperatures; however, there is no report of cryocrystallography for membrane proteins. In this communication, we report on a successful experiment in which flash-cooled bc1 membrane protein crystals have given rise to sustained diffraction over a 60 hour data collection period at a synchrotron source. Furthermore, we present an improved purification and crystallization protocol yielding crystals readily diffracting out to 3.3 A. These results should greatly aid in the future realization of the molecular structure of the bc1 complex as well as other membrane proteins.

Animals↗
PubMed1995

Resection of hepatocellular carcinoma with diaphragmatic invasion.

Fourteen consecutive patients with hepatocellular carcinoma (HCC) and diaphragmatic invasion found during operation and confirmed histopathologically were studied. They were treated by resection of the liver tumour en bloc with part of the diaphragm. Eight of the 14 patients had a diaphragmatic hump on chest radiography. Preoperative ultrasonography, computed tomography and hepatic angiography were unable to diagnose diaphragmatic invasion in these patients. A control group of 14 closely matched patients were selected from 98 undergoing curative resection for HCC during the period of the study. No evidence of a diaphragmatic hump was found in the control patients (P < 0.001). There was no significant difference in survival, operative morbidity or mortality rates between the two groups. In patients with HCC situated in the dome of the liver, the presence of a diaphragmatic hump on chest radiography strongly suggests invasion of the diaphragm. There was no difference in outcome in patients with or without diaphragmatic invasion; the tumour should be resected if the patient is fit for operation.

Carcinoma, Hepatocellular↗
PubMed1995

Detection and assessment of intracranial aneurysms: value of CT angiography with shaded-surface display.

OBJECTIVE: The purpose of this study was to evaluate the efficacy of circle of Willis CT angiography with shaded-surface display in the detection and assessment of suspected intracranial aneurysms. MATERIALS AND METHODS: Twenty-three patients who had clinical or radiologic suspicion of intracranial aneurysms were studied prospectively with CT angiography and conventional angiography. The images were obtained and interpreted in a double-blind fashion by different radiologists. Conventional angiography, which was the reference standard in this study, diagnosed 17 aneurysms in 15 patients. The usefulness of the two types of images for surgical planning was evaluated by two neurosurgeons. RESULTS: No aneurysms were detected with either technique in eight patients. Both techniques showed 15 aneurysms in 14 patients. The maximum dimensions were less than 3.0 mm in three cases, 3.0-5.0 mm in four cases, and greater than 5.0 mm in eight cases. With conventional angiography used as the reference standard, CT angiography has one false-positive and two false-negative findings, resulting in a sensitivity of 88% (15/17) and a specificity of 89% (8/9). For 12 of 15 aneurysms, CT angiography was rated equal or superior to conventional angiography in depicting all aspects (shape, orientation, neck, and parent vessel) of the aneurysms. CONCLUSION: We were able to detect aneurysms of the circle of Willis as small as 2 mm in size. Using conventional angiography as the reference standard, CT angiography has a sensitivity of 88% and a specificity of 89% for the detection of aneurysms in the circle of Willis. In almost all cases, CT angiography was equal or superior to conventional angiography in characterizing the aneurysms for surgical planning.

Adolescent↗
PubMed1995

[Diagnostic score for acute appendicitis].

Scoring systems seem to be ideal for supporting diagnosis of acute appendicitis because they are non invasive, require no special equipment and can be used in clinical routine. Several scores for appendicitis have been developed with good results in the original publications. Unfortunately these good results could not be reproduced on a German data base. Therefore we developed a new score using multivariate statistics and a quality controlled prospective data base. The score covers 8 variables: tenderness, rebound tenderness, micturition, type of pain, leucocytes, age, relocation of pain, rigidity. Independent evaluation of the score on a Dutch database resulted in a negative appendicectomy rate of 21% and a missing appendicitis rate of 2%. The results are encouraging, so that further testing and clinical application can be recommended.

Acute Disease↗
PubMed1994

Mutations affecting the activity of toxic shock syndrome toxin-1.

Toxic shock syndrome toxin-1 (TSST-1), the potent staphylococcal exoprotein linked to most cases of the toxic shock syndrome, is a V beta-restricted T-cell mitogen (a so-called "superantigen"). TSST-ovine (TSST-O) is a natural variant of TSST-1, and is produced by certain ovine mastitis-associated strains of Staphylococcus aureus. Compared to TSST-1, TSST-O is only weakly mitogenic for leporine or murine splenocytes. It differs from TSST-1 at 7 amino acid residues over its 194 amino acid length. Terminus shuffling between the two proteins has suggested that their C-terminal differences (T69, Y80, E132, and I140 in TSST-1; 169, W80, K132, and T140 in TSST-O) are in part responsible for their discrepant mitogenic properties. In order to explore further the functional consequences of altering TSST-1 at residues 132 and 140, we engineered point mutants of TSST-1 at those positions. The mutant proteins were purified to homogeneity from culture supernants of a nontoxigenic strain of S. aureus using a combination of ultrafiltration, liquid-phase isoelectric focusing, and ion-exchange chromatography. The mutants retained global structural integrity as evidenced by circular dichroism spectroscopy, their preserved resistance to trypsin digestion, and their preserved binding to a neutralizing murine monoclonal antibody. The mutants were then tested for mitogenicity for human T-cells: The mutant I140T was approximately as active as wild-type TSST-1, while the mutant E132D was about 10-fold attenuated. On the other hand, the mutants E132A or E132K were each at least 1000-fold attenuated.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacterial Toxins↗
PubMed1994

Decay-accelerating factor (CD55), a glycosylphosphatidylinositol-anchored complement regulatory protein, is a receptor for several echoviruses.

Echoviruses are human pathogens belonging to the picornavirus family. Decay-accelerating factor (DAF) is a glycosylphosphatidylinositol (GPI)-anchored surface protein that protects cells from lysis by autologous complement. Anti-DAF monoclonal antibodies prevented echovirus 7 attachment to susceptible cells and protected cells from infection. HeLa cells specifically lost the capacity to bind echovirus 7 when treated with phosphatidylinositol-specific phospholipase C, an enzyme that releases GPI-anchored proteins from the cell surface, indicating that the virus receptor, like DAF, is a GPI-anchored protein. Although Chinese hamster ovary cells do not bind echovirus 7, transfectants expressing human DAF bound virus efficiently, and binding was prevented by pretreatment with an anti-DAF monoclonal antibody. Anti-DAF antibodies prevented infection by at least six echovirus serotypes. These results indicate that DAF is the receptor mediating attachment and infection by several echoviruses.

Animals↗
PubMed1994

T cells and natural killer cells regulate human IgG subclass concentrations in SCID mice.

Cytokine regulation of human IgG subclass production is not well understood. Since T cells and natural killer (NK) cells produce IL-4 and/or IFN-gamma, we examined the effect of these cells on human IgG subclass concentration in reconstituted severe combined immunodeficient (SCID) mice. SCID mice receiving only B-cell-enriched splenocyte preparations had significantly decreased IgG concentrations and significantly decreased IgG1/IgG2 ratios compared to mice receiving B cells plus T cells (P = 0.02). IgG2 represented 58% of the total IgG at 28 days in mice receiving B-cell-enriched preparations compared to 19% of the total for the group receiving both B cells and T cells (P = 0.013). The effect of natural killer cells (CD16+) on IgG subclass was also studied in this model. IgG2 concentrations were twofold higher in mice receiving CD16-depleted cells compared to controls (P = 0.004). No significant differences were noted for IgG1, IgG3, or IgG4 subclass concentrations. In conclusion, T cells and natural killer cells influence human IgG subclass regulation in the SCID mouse model. We propose that the regulation of human IgG subclass production can be further examined in the SCID model.

Animals↗
PubMed1994

Experience with the management of ovarian germ cell tumors in Chinese patients.

Treatment results of 37 consecutive patients with primary ovarian germ cell tumors (OGCTs) were analyzed. Thirty-three were referred after initial laparotomy and four were first seen at relapse. Four patients with stage I dysgerminoma and grade 1 immature teratoma were observed after operation without recurrence. There was also no relapse in eight patients with dysgerminoma given postoperative irradiation (whole abdomen, median 30 Gy). Twenty-five patients (3 dysgerminomas, 11 immature teratomas, 9 endodermal sinus tumors, and 2 mixed germ cell tumors) received short-term cis-platinum-based chemotherapy. Six out of eight measurable tumors treated by chemotherapy had complete remission. Complete follow-up information was obtained in 35 out of 37 patients. The 4-year actuarial survival rates of the whole group and those referred immediately after initial surgery were 94.1 and 100%, respectively. cis-Platinum was substituted by carboplatin in eight cases but this did not affect treatment result. Nonetheless, deaths occurred in two of four patients referred at relapse with extensive disease and initially treated with suboptimal regimens. Chemotherapy-induced side effects were common but mostly tolerable and were related to cis-platinum and bleomycin. The results of this series show that cis-platinum-based chemotherapy is so effective that nearly 100% cure can be achieved in OGCTs and suggest that it is important to institute optimal chemotherapy from the start. On the other hand, common side effects of treatment and possible late toxicities make it desirable for future studies to see whether chemotherapy intensity could be reduced in patients with good prognosis.

Adolescent↗
PubMed1994

Determination of tumour vascularity using selective hepatic angiography as compared with intrahepatic-arterial technetium-99m macroaggregated albumin scan in hepatocellular carcinoma.

A total of 51 patients with hepatocellular carcinoma (HCC) were studied to determine the tumour vascularity as determined by selective hepatic angiography (HAG) and by intrahepatic-arterial technetium-99m-labeled macroaggregated albumin (Tc99m-MAA) scan. The tumour vascularity was graded on the HAG films by an interventional radiologist using a scale ranging from 1 (hypovascular) to 4 (extremely hypervascular). The grades of vascularity on HAG were grade 1 in 5 patients, grade 2 in 13 patients, grade 3 in 24 patients and grade 4 in 9 patients. The tumour vascularity on scintigraphy was determined by quantifying the count rates over the tumour and normal liver areas by an analog/digital gamma-camera, and the resultant tumour-to-normal ratio (T/N ratio) gave a quantitative measure of the vascularity. The range of the T/N ratio was 0.9 to 11.1, with a median of 3.7. There was no correlation between the tumour vascularity grading on HAG and the T/N ratio on the Tc99m-MAA scan (Wilcoxon rank test, P = 0.83). Thus, we conclude that HAG cannot reveal the true vascularity nor reflect the T/N ratio in HCC.

Angiography↗
PubMed1994

Cytoreductive surgery for hepatocellular carcinoma.

A prospective study was conducted on 26 patients for cytoreductive surgery of inoperable hepatocellular carcinoma. These patients underwent cytoreduction with liver resection, cryosurgery, microwave tissue coagulation and/or absolute alcohol injection. In-hospital mortality was 7.7%. The symptomatic relief and quality of survival were excellent. The median survival of patients after cytoreduction was 10.0 months and the survival was much better than those of 26 patients matched by sex, age, tumour size, Child-Pugh grading and Karnofsky scores who received systemic chemotherapy during the same period of the study (log rank test, P = 0.0001). There was no statistical difference between the survival curves of those patients who received (19 patients) and those who did not receive (7 patients) additional treatment by chemotherapy or selective internal radiation therapy after cytoreduction. This suggests that the gained survival benefit could have been derived mainly from the cytoreductive surgery rather than the additional treatments.

Adolescent↗
PubMed1994

Treatment of inoperable hepatocellular carcinoma with intrahepatic arterial yttrium-90 microspheres: a phase I and II study.

Eighteen patients with inoperable hepatocellular carcinoma (HCC) were treated with intrahepatic arterial yttrium-90 microspheres. All these patients showed a lung shunting below 15% and a tumour-to-normal ratio higher than 2 as determined by diagnostic technetium-99m macroaggregated albumin (Tc-MAA) gamma scintigraphy. The treatment was given through an arterial port placed during laparotomy. The radiation doses to the liver and tumour were determined intraoperatively with a beta probe and liquid scintillation counting of multiple liver biopsies. The treatment was well tolerated without major complications. In all patients the tumour marker fell to a level which ranged from 41% to 0.2% of the pretreatment level. Tumour regression was found to be dose related. Progressive or static disease occurred in a higher proportion of patients whose tumours received < 120 Gy (P = 0.005). Survival was better in those whose tumours received > 120 Gy (median survival = 55.9 weeks) than those whose tumours received lower doses (median survival = 26.2 weeks). This difference is statistically significant with P = 0.005. We conclude that yttrium-90 microsphere therapy is safe and that tumour response is dose related. A tumour dose of > 120 Gy is recommended.

Adolescent↗