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Biomedical subjects

H M Cheng

Publications and source records attributed to H M Cheng.

At least 109 records · Page 6Linked to original sources

Serum and salivary IgA antibodies against a defined epitope of the Epstein-Barr virus nuclear antigen (EBNA) are elevated in nasopharyngeal carcinoma.

The Epstein-Barr virus nuclear antigen I (EBNA I) is the only latent EBV antigen consistently expressed in malignant tissues of the nasopharynx. A 20-amino-acid synthetic peptide, p107 contains a major epitope of EBNA I. We tested sera from 210 patients with nasopharyngeal carcinoma (NPC) and from 128 normal individuals (NHS) for IgA antibodies to p107 using an enzyme-linked immunosorbent assay (ELISA). Whereas 191/210 (91%) of NPC patients had IgA antibodies to p107, only 17/128 (13.3%) of NHS had such antibodies and only 6/57 (10.5%) of sera from patients with malignancies other than NPC had IgA-p107 reactivity. Thirty-nine salivary samples from 46 NPC patients (84.8%) also contained IgA-p107 antibodies whereas only 3/42 (7.1%) of normal saliva samples were IgA-p107 positive. The results suggest that IgA antibodies to EBNA I may become a useful, easily measurable, marker for NPC.

Adult↗

Metabolic analysis of reepithelializing rabbit cornea using phosphorus-31 nuclear magnetic resonance spectroscopy.

To investigate metabolic differences between the central and peripheral cornea the latter including the limbal area, corneas were dissected and examined using phosphorus-31 (31P) nuclear magnetic resonance spectroscopy. Since most 31P signals originate from the epithelium, 31P spectra of the cornea primarily represent the metabolic state of the epithelium. The spectra of the peripheral cornea showed all phosphorus resonances detected in the whole cornea; in contrast, the central cornea showed no phosphocreatine and glycerophosrylethanolamine, and only low levels of ATP. These results indicate that there is a higher metabolic activity in the peripheral epithelium, especially in the limbal area, than in the central epithelium. To evaluate the metabolic state of corneal epithelium during regeneration, we also examined corneas reepithelializing after 7 mm of central epithelial tissue had been removed by mechanical scraping. Rabbits were killed 24 and 48 h after scraping. The reepithelializing corneas clearly showed an increase in ATP, phosphocreatine, and sugar phosphates with time, although phosphorylcholine remained depressed. These findings suggest that the reepithelializing cornea has an elevated level of energy production and that it may have reached a higher steady state, thereby indicating accelerated metabolism of the epithelium during regeneration.

Animals↗

Metabolic studies of galactosemic cataract.

Experimental diabetic and galactosemic animal models are widely used to study diabetes-induced complications. Galactose feeding can rapidly produce cataract, retinopathy and nephropathy; it is therefore favored over the diabetic model. Although the common feature for both models is the activation of aldose reductase, there are substantial differences between the two--not only does the rate of cataract progression differ but the metabolic patterns are far more complex than for polyol production alone. We here present the result of a comparison between diabetic and galactosemic lenses and show the differences in phosphorus and aldose metabolism, cell integrity and osmotic environment.

Animals↗

Bacterial immunity and immunogenesis of normal human salivary IgA and serum IgG2 antiphospholipid autoantibody: a link?

The anti-phospholipid antibody (aPL) in 26 heat-inactivated normal human sera (NHS) was tested for IgG subclass in ELISA. The specific antibody in NHS included all four IgG antibody subclasses, as well as IgA. The incidence of IgG subclasses ranged from 50% (13/26) for IgG1 to 92% (24/26) for IgG2. Specific IgA anti-phospholipid antibody (aPL) was detected by ELISA in 38% (28/73) of normal human saliva. The salivary IgA aPL bound preferentially to anionic phospholipids including cardiolipin, phosphatidylserine and phosphatidic acid but not to phosphatidylcholine or sphingomyelin. Unlike aPL in normal human sera, aPL in saliva was predominantly not associated with the previously described heat-labile inhibitor of aPL. This may indicate a role of salivary IgA aPL in local immunity by binding to cross-reactive bacterial cell surface components including phospholipids.

Antibody Specificity↗

Anti-phospholipid antibody isotypes in normal human sera.

Heat-sensitive serum masking cofactor(s) of antiphospholipid antibody (aPL) in normal human sera (NHS) are specifically inactivated at 56 degrees C. The degree of binding in ELISA by unmasked aPL in NHS was equivalent to that in non-heated, aPL-reactive autoimmune SLE sera. Previously "negative" SLE sera also reacted equally strongly in the aPL ELISA when similarly heat-inactivated. Isotype studies by ELISA of the heat-potentiated aPL in 36 NHS revealed the presence of specific IgG (34/36), IgM (11/36) and IgA (24/36) aPL antibodies. 11/36 (31%) NHS had all three aPL isotypes while 13/36 (36%) had both IgG and IgA antibodies to phospholipid.

Cardiolipins↗

ATP, ADP, and magnesium mixed solutions: in vitro 31P NMR characterization and in vivo application to lens.

The difference between the 31P NMR resonance position of the beta-peak of ADP and the gamma-peak of ATP at a fixed monovalent ion concentration level and pH is shown in vitro to depend only on free magnesium concentration. This difference can vary by more than 1 ppm depending on the pH of the solution and free magnesium concentration. Using 31P NMR and ion-selective electrodes we have constructed experimental curves to show how calibration of experimental results can be achieved. Theoretical calculations can be used to predict the functional dependence of the chemical shift difference on the above parameters. However, using the reported ion dissociation constants the fit was not exact and the discrepancy increased at higher pH values. To demonstrate that this technique can yield valid in vivo results and to analyze a previously unreported system where free magnesium levels vary, sample spectra from lenses and enucleated eyes are given. The data show a disagreement between the present methods and the more conventional ATP method. The reason for this difference is not known although some possible reasons are suggested.

Adenosine Diphosphate↗

Fanconi syndrome: report of a case.

A 59-year-old female patient was admitted because of muscle weakness in all four limbs for a period of 5 days. She had been found to have Graves' disease 4 years ago previous to this, and had received a subtotal thyroidectomy 1 year later. Hypothyroidism supervened and she had been receiving levothyroxine replacement in recent years. She also had non-insulin-dependent diabetes, which was controlled with diet only. During the 5 days prior to admission, she developed muscle weakness which finally worsened to complete paralysis of all four limbs. Physical examination showed tenderness and weakness of the extremity muscles. Abnormal laboratory data included serum K, 1.6 mEq/L; P, 1.2 mg/dl; uric acid, 1.6 mg/dl; fasting glucose, 267 mg/dl; T3, 36.65 ng/dl; T4, 4.0 micrograms/dl; TSH, 5.35 mu u/ml; free T4, 0.57 ng/dl; and metabolic acidosis with pH, 7.298; PCO2, 27.0 mmHg; and HCO3, 12.8 mEq/L. An EKG showed a prominent U wave, and urinalysis revealed renal glucosuria and massive aminoaciduria. An oral sodium bicarbonate loading test showed an increasing loss of bicarbonate through the urine, while the plasma bicarbonate level was elevated. Clinical manifestations improved after the administration of sodium bicarbonate, potassium chloride and neutral phosphate.

Autoimmune Diseases↗

Heat inactivation of serum potentiates anti-cardiolipin antibody binding in ELISA.

Heat treatment of sera at 56 degrees C for 30 min results in positive ELISA reactions for anti-cardiolipin antibody (aCL) in sera that had undetectable or low levels of aCL before heat inactivation. The positive, potentiated reactivity of the heated sera in the aCL ELISA could be inhibited with the cardiolipin antigen and was abolished by prior IgG depletion using staphylococcal protein A. The heat-potentiating effect of aCL binding in ELISA was evident in both normal human sera and clinical sera including sera from patients with systemic lupus erythematosus and syphilis.

Cardiolipins↗

Improved sensitivity of detection by avidin-biotin complex (ABC) immunocytochemistry in Epstein-Barr virus serology.

Serum antibodies against Epstein-Barr virus (EBV)-determined antigens have traditionally been titrated by the indirect immunofluorescence (IIF) technique. The avidin-biotin complex (ABC) immunocytochemical technique was used to determine the serum levels of IgA against EBV viral capsid antigen (IgA/VCA) and IgA against EBV early antigen (IgA/EA) in sera of 106 nasopharyngeal carcinoma (NPC) patients prior to treatment and 100 normal individuals. The sensitivity of the ABC technique is enhanced by an amplification of the antigen-antibody reaction, which involves the binding of the enzyme-linked ABC to the second biotinylated antibody. There was a good correlation (r = 0.9988) between ABC and IIF-determined IgA/VCA-positive titres, with the ABC technique being more sensitive than IIF in the detection of IgA/VCA in NPC sera: 94% (99/106) and 76% (80/106), respectively. The frequency of IgA/EA reactivity in NPC sera was also markedly increased by immunodetection with the ABC technique as compared with IIF technique: 63% (69/106) and 28% (30/106) respectively. Both the immunocytochemical techniques were equally specific in discriminating between elevated serum titres of IgA/VCA and IgA/EA in NPC sera from normal human sera.

Antibodies, Viral↗

Lens redox fluorometry: pyridine nucleotide fluorescence and analysis of diabetic lens.

We performed both ex vivo and in vivo fluorometric analyses of pyridine nucleotides (PN) in rabbit and rat lenses. Rabbit lens PN fluorescence (99% NADH) was found to have an excitation maximum at 366 nm and an emission maximum at 462 nm (366:462). The only other fluorescent chromophore in that region of the spectrum has excitation and emission peaks at 328 and 460 nm, respectively. Anaerobic glycolysis in the lens was stimulated by KCN, a known inhibitor of mitochondrial respiration, after which a time-study of fluorescence intensities was performed. Over the course of a 3.5 hr period following treatment with KCN, the PN signal showed a statistically significant increase relative to that in the control lenses (those treated with KCl). while the 328:460 signal (which may be due to some protein involved in energy transfer with the PN) had a significantly greater decrease. We also found that fluorescence intensity of NADH in solution is linearly proportional to physiologic-range concentration. Moreover, there was a close correlation between fluorescence intensity of rat lens PN as measured on a specular microscope-coupled redox fluorometer capable of in vivo use, and the lens PN levels as determined by the analytical cycling assay technique. This fluorometer was then employed to assess the redox state in rats with streptozotocin-induced diabetes. The normalized ratio of PN to flavoproteins (Fp) in the lens epithelium increased from 0.96 +/- 0.12 in the normal state to 1.48 +/- 0.30 2 weeks after diabetes induction. In contrast, the ratio in the diabetic lens treated with an aldose reductase inhibitor, sorbinil, did not increase. The increase in the PN:Fp ratio therefore reflects activation of the polyol pathway and its associated metabolic activities, which results in an increase in the NADH:NAD ratio in the diabetic rat lenses. Our results indicate that the non-invasive, real-time method of redox fluorometry may be useful in the early detection and evaluation of cataracts and other disorders in lens metabolism, long before opacities occur. It can be used to monitor the disease process and evaluate the efficacy of such drugs as aldose reductase inhibitors on a biochemical level.

Animals↗

Polyol pathway activity in streptozotocin-diabetic rat lens.

We have examined polyol pathway kinetics in the lenses of rats made diabetic with streptozotocin. At up to 11 days after diabetes induction, the lenses were isolated and subjected to 'pulse-chase' studies: the lenses were incubated with [13C]glucose and lens metabolism followed by [13C]nuclear magnetic resonance (NMR) spectroscopy. Proton NMR spectroscopy was also performed to measure the hexose monophosphate shunt (HMPS) activity. The results showed that (1) the activity of aldose reductase increased initially and decreased after 11 days of diabetes; (2) the fructose pool increased initially but started to decline after 3 days; (3) the HMPS activity increased nearly 40% immediately after diabetes induction; and (4) the turnover rates of glucose, alpha-glycerophosphate (GP), lactate, sorbitol, and fructose were 80.8 +/- 2.6, 10.1 +/- 1.4, 47.7 +/- 3.7, 7.9 +/- 0.9 and 5.2 +/- 2.2 nmol hr-1 lens-1 (34 mg wet weight lens-1), respectively. Up to 35% of lactate appeared to derive from the polyol pathway. Further, GP was rapidly metabolized, although its fate is currently unknown. These results reveal a far more complex pattern of glucose metabolism in the diabetic lens than that in lenses incubated in high glucose.

Animals↗

Heat-labile serum inhibitor of anti-phospholipid antibody binding in ELISA.

Normal human sera (NHS), heat-inactivated at 56 degrees C for 30 min, demonstrated positive ELISA reactions for anti-cardiolipin (aCL) antibodies. The heat-induced reactivity in ELISA was inhibitable by the cardiolipin antigen and was abolished by prior IgG depletion of the heated NHS with a protein A preparation. The heat-potentiated aCL also cross-reacted selectively with phosphatidic acid and phosphatidylserine, but not with phosphatidylcholine or phosphatidylethanolamine.

Biological Factors↗

31P NMR studies of excised gray and white calf brain.

1. 31P NMR examination of isolated calf gray and white matter reveals that white matter contains higher levels of the phosphodiester glycerolphosphoryl choline (GPC) than gray. 2. It is suggested that GPC may play a role in maintaining the level of phospholipids present by inhibition of phospholipases. 3. The spectra also reveal a skewed peak whose maximum is at -11 ppm which is inferred to arise from myelin-like structures. 4. The results show that phosphorus spectra from the brain must be carefully considered whether they arise from the same type tissue or represent a mixed sample since variation in results may represent anatomy as well as physiology.

Animals↗

Hard contact lens-induced metabolic changes in rabbit corneas.

The biochemistry of contact lens-cornea interaction is not well understood, although previous studies have suggested that corneal metabolic changes may be the underlying factor in morphological alterations. Using a rabbit model, this interaction has been examined with 31P nuclear magnetic resonance (NMR) spectroscopy, which detects signals principally from the epithelium. The examination was supplemented with electron microscopy and histochemistry. Polymethylmethacrylate lenses caused reversible changes, including activation of anaerobic glycolysis and disturbance of membrane metabolite levels. These changes were far more severe than those occurring during prolonged eye closure. There appears to be an association between cellular deterioration and loss of membrane metabolites. On the other hand, oxygen-permeable silicone lenses allowed maintenance of nearly normal metabolic patterns. These results show multifaceted corneal response to hard contact lens wear.

Animals↗

Cornea-soft contact lens interaction: a nuclear magnetic resonance study.

Because 31P NMR (phosphorus-31 nuclear magnetic resonance) spectroscopy can detect phosphorus metabolites and more than 90% of them reside in corneal epithelium, we applied this technique to the examination of the metabolic response of the cornea (epithelium) to soft contact lens (CL) wear. Our results showed that 4 h of soft CL (59% water) wear caused a 50% loss of ATP, and both ATP and phosphocreatine (PCr) decreased drastically if the eyes were closed. Topical application of 5.5 mM glucose to CL-fitted eyes restored the 31P profile to nearly normal. Possible approaches for maintaining corneal health are discussed.

Adenosine Triphosphate↗