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Biomedical subjects

C W Lam

Publications and source records attributed to C W Lam.

At least 163 records · Page 9Linked to original sources

High-performance liquid affinity chromatography for the purification of immunoglobulin A from human serum using jacalin.

A high-performance liquid affinity chromatographic method for the purification of serum immunoglobulin A (IgA) using a jacalin column is described. The automated procedure takes about 2 with minimal manipulation. The yields of the isolated IgA and of its IgG and IgM contamination were studied by enzyme-linked immunosorbent assay (ELISA) of 30 sera. Purity was assured by immunoelectrophoresis. The ratio of IgA1 to total IgA was unchanged after purification, as verified by ELISA. The results showed that greater than 90% IgA could be recovered with less than 0.5% total IgG and greater than 2.0% total IgM remaining in the fractions containing purified IgA.

Chromatography, Affinity↗

Mechanism of transport and distribution of organic solvents in blood.

Little is known about the mechanism of transport and distribution of volatile organic compounds in blood. Studies were conducted on five typical organic solvents to investigate how these compounds are transported and distributed in blood. Groups of four to five rats were exposed for 2 hr to 500 ppm of n-hexane, toluene, chloroform, methyl isobutyl ketone (MIBK), or diethyl ether vapor; 94, 66, 90, 51, or 49%, respectively, of these solvents in the blood were found in the red blood cells (RBCs). Very similar results were obtained in vitro when aqueous solutions of these solvents were added to rat blood. In vitro studies were also conducted on human blood with these solvents; 66, 43, 65, 49, or 46%, respectively, of the added solvent was taken up by the RBCs. These results indicate that RBCs from humans and rats exhibited substantial differences in affinity for the three more hydrophobic solvents studied. When solutions of these solvents were added to human plasma and RBC samples, large fractions (51-96%) of the solvents were recovered from ammonium sulfate-precipitated plasma proteins and hemoglobin. Smaller fractions were recovered from plasma water and red cell water. Less than 10% of each of the added solvents in RBC samples was found in the red cell membrane ghosts. These results indicate that RBCs play an important role in the uptake and transport of these solvents. Proteins, chiefly hemoglobin, are the major carriers of these compounds in blood. It can be inferred from the results of the present study that volatile lipophilic organic solvents are probably taken up by the hydrophobic sites of blood proteins.

Administration, Inhalation↗

The influence of hyperglycemia and diabetes mellitus on immediate and 3-month morbidity and mortality after acute stroke.

Fasting serum glucose, glycosylated hemoglobin, and fructosamine concentrations were determined in 304 consecutive subjects admitted with acute stroke, within 48 hours of ictus. Based on the medical history and these results, subjects were divided into known diabetics, newly diagnosed diabetics, subjects with stress hyperglycemia, and nondiabetics. The type of stroke was classified as lacunar infarct, cerebral infarct, or intracerebral hemorrhage, based on clinical examination by a neurologist and computed tomographic brain scan and/or autopsy. Immediate and 3-month outcomes were examined in relation to (1) fasting glucose, glycosylated hemoglobin, and fructosamine levels by stroke subtypes, and (2) glucose tolerance categories by stroke subtypes. A high fasting glucose level was associated with an increased mortality, but this was observed only among patients with intracerebral hemorrhage. Patients with stress hyperglycemia, but not diabetics, had increased mortality. In spite of having similar glucose concentrations to those patients with stress hyperglycemia, diabetics did not have a worse outcome compared with nondiabetics. It is concluded that the association between glucose concentration and outcome is a reflection of stress relating to stroke severity, rather than a direct harmful effect of glucose on damaged neurons.

Acute Disease↗

A case control study of some hematological and biochemical variables in acute stroke and their prognostic value.

Hematocrit, total white cell and platelet counts, plasma urea and creatinine concentrations were estimated in 291 patients (mean age 70 +/- 12 years) admitted consecutively over a 6-month period to a district general hospital with acute stroke. Urinary albumin/creatinine ratio was also determined in a subgroup of patients during a follow-up visit 3 months after ictus. Results were compared with those from age- and sex-matched community controls. Total white cell count was elevated in all types of stroke compared with values from control subjects. Platelet count, plasma urea and creatinine concentrations were higher and the plasma albumin/globulin ratio was lower among patients with ischemic stroke. Elevated total white cell count, urea and creatinine, which were of prognostic significance, were found to be associated with the severity of stroke as indicated by the Glasgow coma score, and did not have any independent prognostic value. Elevated hematocrit was not shown to be a risk factor and did not have any prognostic significance. A low plasma albumin/globulin ratio, an index of plasma viscosity, was a possible risk factor for ischemic strokes. Urinary albumin/creatinine ratios in the highest quintile increased the risk of stroke 13-fold.

Aged↗

Acute and long-term changes in serum lipids after acute stroke.

We studied serum lipid profiles in 171 patients less than or equal to 48 hours after the onset of acute stroke and 3 months later. The 83 patients suffering cerebral infarction had significantly higher serum concentrations of total cholesterol, low density lipoprotein-cholesterol, and apolipoprotein B and significantly lower serum concentrations of triglycerides and lipoprotein (a) less than or equal to 48 hours after ictus than 3 months later. The lipid profiles of the 53 patients suffering lacunar infarction were similar on both occasions, the only significant differences being higher total cholesterol and low density lipoprotein-cholesterol concentrations less than or equal to 48 hours after ictus. No significant changes were observed among the 35 patients suffering cerebral hemorrhage apart from a significantly higher concentration of high density lipoprotein3-cholesterol less than or equal to 48 hours after ictus. Our study, with many patients classified according to stroke subtype, gives results different from those of previous studies with much fewer patients. We conclude that in studies of serum lipid and lipoprotein concentrations as risk factors for cerebral infarction, comparing values obtained less than or equal to 48 hours after admission with control values may incorrectly identify certain lipid fractions as risk factors.

Aged↗

Zinc and birth weight in uncomplicated pregnancies.

The relationship between zinc status and birth weight in uncomplicated pregnancies was studied in 59 nulliparous and 27 multiparous Chinese women with singleton pregnancies, delivered at term. Maternal and umbilical cord blood, maternal pubic hair and a segment of the umbilical cord were collected at delivery for analysis. The zinc concentration was significantly higher (p less than 0.0001) in the plasma of newborn infants (13.9 +/- 2.8 mumol/l) than in maternal plasma (9.3 +/- 2.2 mumol/l), while the converse was found in tissue (60.8 +/- 17.0 vs 208.1 +/- 96.2 micrograms/g, p less than 0.0001). There was no significant correlation between birth weight and either maternal or newborn plasma or tissue zinc concentrations. Parity had no significant effect on maternal or newborn zinc concentrations in tissue and plasma.

Birth Weight↗

Light-chain ratios of immunoglobulins G, A, and M determined by enzyme immunoassay.

We describe an enzyme-linked immunosorbent assay for determination of light-chain ratios for IgG, IgA, and IgM in serum. A commercial serum with known overall kappa and lambda concentrations was used as standard. To capture the respective immunoglobulins, we used antibodies to gamma, lambda and mu chains coated onto microtiter plates. Peroxidase-conjugated anti-kappa and anti-lambda chain antisera were reacted with light chains on the captured immunoglobulins, and the amount of enzyme bound was monitored with o-phenylenediamine and urea-hydrogen peroxide as substrates. Calculation of absorbance ratios allowed determination of kappa and lambda chain concentrations of individual immunoglobulins in the standard and samples. Within-run and between-run CVs (n = 25) ranged from 5.9% to 13.0% for "high," "normal," and "low" kappa/lambda ratios for IgG, IgA, and IgM. The thoroughness of light-chain detection, expressed as, e.g., (IgA kappa + IgA lambda)/(total IgA), for 150 sera was 91-110%. The detection limit was 1 microgram/L. Reference intervals (mean +/- SD) for kappa/lambda ratios in sera from 100 apparently healthy adults were 2.34 +/- 0.80 for IgG, 1.59 +/- 0.40 for IgA, and 1.86 +/- 0.76 for IgM.

Adult↗

Effect of ciclosporin on lymphocyte subpopulations and immunoglobulin production in IgA nephropathy.

The effect of ciclosporin on the cellular immunoregulation was examined in 19 patients with IgA nephropathy. The patients were randomly divided into two groups: 9 patients receiving oral ciclosporin (5 mg/kg/day) for 12 weeks and 10 patients receiving placebo and acting as controls. T lymphocyte subpopulations were determined using OKT monoclonal antibodies. The functional capability of lymphocytes was assessed by in vitro immunoglobulin synthesis of cultured peripheral mononuclear cells, thymidine uptake by cultured lymphocytes, and t lymphocyte activation with expression of interleukin-2 receptors. A fall of in vitro IgA production by cultured lymphocytes (p less than 0.05), a reduction of thymidine uptake by Staphylococcus aureus Cowan-stimulated cultured lymphocytes (p less than 0.05), and a reduction of activated lymphocytes expressing interleukin-2 receptor (p less than 0.05) were observed in patients after 12 weeks of ciclosporin therapy. The percentages of OKT4 and OKT8 lymphocytes and OKT4/8 ratios were not altered with therapy. A simultaneous reduction of proteinuria and a transient impairment of renal function were observed. Similar changes in cellular immune parameters and clinical response were not observed in the controls. Our study suggests ciclosporin could modulate the cellular immunity in IgA nephropathy.

Cyclosporins↗

Enzyme-linked immunosorbent assay for alpha-fetoprotein in human serum.

A simple and inexpensive enzyme-linked immunosorbent assay for determination of serum alpha-fetoprotein concentration, with results available within 4 h, is described. The assay has a working range of 2-100 ng/ml, with within- and between-batch coefficients of variation (n = 24) at different concentrations between 8.9 and 12.7%. Results on 80 patients' sera (AFP concentrations 0-175 ng/ml) correlate well with those by radioimmunoassay. Turn around time (4 h) is shorter than with similar immunoassays currently available.

Carcinoma, Hepatocellular↗

Direct determination of lead in urine by flameless atomic absorption spectrophotometry with platform atomisation.

A direct method of urinary lead assay by flameless atomic absorption spectrophotometry is described for routine use in the clinical laboratory. It employs a simple sample pre-treatment procedure preceding automated analysis. Reproducibility, linearity and recovery were satisfactory without the need of standard addition. Mean and reference intervals of normal urine were similar to values reported in the literature.

Humans↗

Enzyme-linked immunosorbent assay for free thyroxin in human serum.

We describe a simple enzyme-linked immunosorbent assay (ELISA) in which microtiter plates are used for determining the free thyroxin concentration (FT4) in serum. Only commercially available chemicals and reagents are needed, including the antiserum. The working range is 1 to 60 ng/L. Turnaround time is about 4 h. Within-run coefficients of variation (CV) for FT4 concentrations of 9.0, 19.0, and 35.0 ng/L were less than 5%; between-run CVs were 10% to 11%. Results by a radioimmunoassay method for 150 sera correlated well (r = 0.922).

Enzyme-Linked Immunosorbent Assay↗

Light chain composition of IgA in IgA nephropathy.

The immunochemical nature of the light chains of the mesangial IgA deposits were studied in 13 IgA nephritic patients with only IgA isotype in their renal biopsies. Direct immunofluorescence (IF) using rabbit and goat antihuman monospecific antibodies demonstrated lambda light chain IF in all 13 biopsies (100%). Kappa light chain IF was detected in ten renal biopsies by the rabbit antihuman antisera and in eight renal biopsies by the goat antihuman antisera. Monoclonal antibodies studies showed lambda IF in all 13 biopsies and kappa IF in eight biopsies. Twelve biopsies showed a predominance of lambda light chain IF staining in the mesangial deposits. Identical IF patterns were observed when double IF technique with rhodamine-conjugated antihuman alpha heavy chain antiserum and fluorescein-conjugated antihuman light chain antisera were employed, and these findings confirmed that the light chains IF arose directly from the IgA deposits. These IgA nephritic patients had a significantly higher serum concentration of lambda light chain-IgA and a distinct alteration of kappa/lambda ratio of IgA compared with healthy controls. Our results support our preliminary observation that mesangial IgA deposits in IgA nephropathy consist mostly of IgA with lambda light chain.

Fluorescent Antibody Technique↗

Predominant synthesis of IgA with lambda light chain in IgA nephropathy.

The nature of the light chains in mesangial IgA deposits and serum IgA was studied in patients with IgA nephropathy. Immunofluorescence (IF) studies using murine monoclonal antibodies, rabbit and goat anti-human monospecific antisera were performed in kidney sections from 15 IgA nephritic patients with only IgA isotype detected in the renal biopsy. Lambda light chain IF was demonstrated in all biopsy specimens and kappa light chain IF in 11 renal biopsy specimens. The majority of renal biopsies showed a predominance of lambda light chain IF staining in the mesangial deposits. The concentration of individual immunoglobulins and their light chain fractions, and the kappa/lambda ratio were determined in the serum and the supernate from peripheral blood mononuclear cells culture of 30 IgA nephritic patients and 30 age-matched healthy controls. The IgA nephritic patients had a higher serum concentration of total IgA (P less than 0.001) and a significantly lower IgA kappa/lambda ratio (P less than 0.001) compared with the controls. The kappa/lambda ratio of supernatant IgA from IgA nephritic patients (N = 20) was also significantly lower than that of the normal subjects (N = 14), both in the unstimulated (P less than 0.01) and pokeweed mitogen stimulated, peripheral blood mononuclear-cell culture (P less than 0.05). Our results showed that patients with primary IgA nephropathy displayed a unique immunologic response characterized by a predominance of IgA with lambda light chain in circulation.

Cells, Cultured↗