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Biomedical subjects

C M Tang

Publications and source records attributed to C M Tang.

At least 55 records · Page 3Linked to original sources
PubMed1996

p53 mutational spectrum of esophageal carcinomas from five different geographical locales in China.

A mutational spectrum for exons 5-8 of the p53 tumor suppressor gene in esophageal carcinomas in mainland China and Hong Kong was established. This study involved 209 squamous cell carcinoma specimens obtained from five different geographical locales in China: Zhengzhou, Taiyuan, Shantou, Guangzhou, and Hong Kong. Zhengzhou and Shantou were high-incidence regions for esophageal cancer, whereas the other three regions had low or intermediate incidence of the disease. Analysis by single-strand conformation polymorphism and DNA sequencing showed that 87 specimens (41.6%) contained mutations in exons 5-8 of the p53 gene compared to 163 cases (78%) that had accumulation or aberrant expression of the protein, as detected by immunohistochemical staining. Point mutations accounted for 80.4% (87/107) of all genetic changes. The specimens from northern China exhibited fewer p53 gene aberrations and a more even distribution of mutations in exons 5-8 compared to those from southern China in which 60% of all mutations were found in exon 5. A major hot spot was found at codon 176 in exon 5, where 41 samples from Shantou, Guangzhou, and Hong Kong had a G-->T transversion. It is likely that among southern Chinese this codon is susceptible to mutagenesis by carcinogens. Codons 175, 203, 245, 250, 273, and 282 were also shown to be mutational hot spots, with three or more mutations observed at each site. The p53 mutational data obtained in this study showed that Chinese esophageal carcinomas are often associated with some unique genetic alterations, which may be attributed to specific dietary or environmental carcinogens that affect the Chinese but not Caucasians.

Adult↗
PubMed1994

Dithiothreitol homogenization of prefixed sputum for lung cancer detection.

The technique of chemical homogenization of sputum for cancer detection was revisited. The mucolytic agent dithiothreitol (DTT) had been used by the authors on fresh specimens. In this study, its effectiveness in homogenizing prefixed sputum was investigated. Fifty-seven positive samples were examined: 28 were prefixed with 2% carbowax in 60% ethanol and 29 in 3% carbowax in 60% ethanol. Each specimen was divided equally into three parts and homogenized immediately, 3 and 7 days later, respectively, with 0.2% (0.013 mol/L) DTT in the respective prefixative. Five samples were prefixed for 4 wk before homogenization. The homogenization time varied from 30 min to 48 hr. The cellular morphology was compared to directly smeared controls from the same samples. It was found that the cellular morphology was well preserved and best with the 3% carbowax in 60% ethanol. The specimen could be prefixed for up to 4 wk before homogenization and the homogenization process could last from 30 min to 48 hr without any damaging effect on morphology. Furthermore, screening was made easy with the mucus lysed and its obscuring effect removed and the cells concentrated and evenly distributed. The method was versatile and could be of value in enhancing the detection rate of cancer in sputum.

Adenocarcinoma↗
PubMed1994

Molecular genetics of Aspergillus pathogenicity.

Aspergillus fumigatus is the most frequent cause of Invasive Pulmonary Aspergillosis (IPA), a life-threatening disease of immunosuppressed patients. In addition to a number of general physiological attributes of this fungus, it has been suggested that extracellular elastase and toxins might facilitate its growth in lung tissue. We have investigated the roles of two extracellular proteins, an alkaline protease with elastase activity (AFAlp), and the ribotoxin restrictocin in murine models of IPA. Gene disruption was used to create stable null mutant strains of the fungus lacking one or other protein, and their virulence and histopathological features were compared with an isogenic parental strain in steroid-treated and neutropenic mice. We have been unable to demonstrate any significant differences between the three strains, which shows that, considered independently, these proteins are not important virulence determinants. We are also interested in identifying fungal-specific gene products involved in general metabolism and which are required for growth in the lung, because these could represent new targets for antifungal drugs. For this work a model of murine IPA involving Aspergillus nidulans was established, to take advantage of the many well characterised mutations affecting metabolic pathways. Pathogenicity tests with strains carrying one of two auxotrophic mutations, lysA2 and pabaA1, have shown while lysine biosynthesis is not essential for the fungus to cause pulmonary disease, biosynthesis of p-aminobenzoic acid is essential. We are now in the process of cloning the A. fumigatus pabaA homologue to determine its function and whether this gene is required for growth of the clinically important species in the lung.

4-Aminobenzoic Acid↗
PubMed1994

Molecular epidemiological study of invasive pulmonary aspergillosis in a renal transplantation unit.

The molecular epidemiology of an outbreak of invasive pulmonary aspergillosis which occurred on a renal transplantation unit was investigated. Restriction fragment length polymorphism and random amplified polymorphic DNA analyses were used to characterise the isolates of Aspergillus fumigatus from the patients (2 isolates from each of 2 patients) and from the environment (11 isolates). While the isolates from the environment could be readily distinguished from each other by both methods, each patient appeared to be infected with a single strain. These findings lend support to the suggestion that certain isolates of Aspergillus fumigatus are more pathogenic than others.

Aspergillosis↗
PubMed1994

Saturation of postsynaptic glutamate receptors after quantal release of transmitter.

Miniature excitatory postsynaptic currents (mEPSCs) were elicited from small numbers of release sites after brief microperfusion of Ba2+ and K+ onto proximal dendritic processes of hippocampal neurons in culture. Temporal summation of closely timed mEPSCs deviated significantly from linearity. The number of instances of closely timed mEPSCs that were also closely matched in terms of peak amplitudes was significantly greater than that expected by chance. Amplitude pairing became statistically more significant after prolongation of mEPSC duration and inhibition of glutamate receptor desensitization with cyclothiazide. These results are best explained by postsynaptic receptors that approach saturation after quantal release of transmitter.

Animals↗
PubMed1994

Expression of non-NMDA glutamate receptor channel genes by clonal human neurons.

Treatment of the human teratocarcinoma line NTera2/c1.D1 (NT2) with retinoic acid induces terminal neuronal differentiation. In a previous study, we found that the neurons obtained in this way express functional N-methyl-D-aspartate (NMDA) and non-NMDA glutamate receptor channels. We now show by reverse transcriptase-polymerase chain reaction and Southern blotting that these neurons transcribe each of the nine known non-NMDA glutamate receptor genes (GluR1-7, Ka-1, and Ka-2) and that four of these genes (GluR2, GluR6, GluR7, and Ka-1) are also transcribed by undifferentiated NT2 cells. Patch clamp studies demonstrate that individual non-NMDA glutamate receptor channels are readily isolated from NT2-derived neurons and that these channels are potently modulated by the desensitization blocker cyclothiazide. NT2-derived neurons are susceptible to kainate excitotoxicity but are not injured by prolonged exposure to alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate. We expect that the NT2-derived human neuronal culture system will facilitate studies of human neuronal non-NMDA glutamate receptor channels and of the pathophysiology of neuronal excitotoxicity.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1994

Virulence of Aspergillus fumigatus double mutants lacking restriction and an alkaline protease in a low-dose model of invasive pulmonary aspergillosis.

To investigate the pathogenicity of Aspergillus fumigatus mutants lacking putative virulence factors, we have developed a new murine model of invasive pulmonary aspergillosis based on neutropenia, the major factor predisposing patients to this infection. Mice were treated with cyclophosphamide and inoculated by the intranasal route with 5 x 10(3) conidia, a significant reduction from inoculum levels used in previous models. Evidence for the production of the extracellular alkaline protease (Alp) in lung tissue was obtained by using a fungal transformant harboring an alp::lacZ reporter gene fusion. The pathogenicities of single mutant strains lacking either Alp or the ribotoxin restrictocin and of a double mutant strain lacking both proteins were assessed in this infection model. There were no significant differences between the mutant and the wild-type strains in terms of mortality or histological-features. Inoculations with mixtures of conidia showed that the double mutant strain is slightly less virulent than the wild-type strain. We conclude that Alp and restrictocin are not important virulence determinants in pulmonary infection.

Allergens↗
PubMed1994

Virulence studies of Aspergillus nidulans mutants requiring lysine or p-aminobenzoic acid in invasive pulmonary aspergillosis.

To identify steps in fungal intermediary metabolism required by Aspergillus spp. during invasive pulmonary aspergillosis, we have developed murine models involving Aspergillus nidulans as the inoculum. The advantages of using A. nidulans over Aspergillus fumigatus or Aspergillus flavus, which are the most common agents of clinical disease, are the well-understood genetics of A. nidulans and a large range of mutants of this species which are affected in a variety of metabolic pathways. Comparison of the virulence of A. nidulans strains carrying mutations which block the biosynthesis of lysine (lysA2) and p-aminobenzoic acid (pabaA1) shows that lysA2 strains have reduced virulence while pabaA1 strains are entirely nonpathogenic. The pathogenicity of pabaA1 strains can be restored by supplementing the drinking water of animals with p-aminobenzoic acid. The results indicate that the availability of lysine in the lung is limited, and p-aminobenzoic acid is probably not available at all. Thus, models of invasive pulmonary aspergillosis involving A. nidulans can be used to identify metabolic pathways that may be essential for the pathogenicity of A. fumigatus, the predominant pathogenic species, suggesting potential new targets for antifungal therapy.

4-Aminobenzoic Acid↗
PubMed1993

Inducible expression of neuronal glutamate receptor channels in the NT2 human cell line.

Glutamate receptor (GluR) channels are responsible for a number of fundamental properties of the mammalian central nervous system, including nearly all excitatory synaptic transmission, synaptic plasticity, and excitotoxin-mediated neuronal death. Although many human and rodent neuroblast cell lines are available, none has been directly shown to express GluR channels. We report here that cells from the human teratocarcinoma line NT2 are induced by retinoic acid to express neuronal N-methyl-D-aspartate (NMDA) and non-NMDA GluR channels concomitant with their terminal differentiation into neuron-like cells. The molecular and physiologic characteristics of these human GluR channels are nearly identical to those in central nervous system neurons, as demonstrated by PCR and patch clamp recordings, and the cells demonstrate glutamate-induced neurotoxicity.

2-Amino-5-phosphonovalerate↗
PubMed1993

Electrophysiologic and molecular properties of cultured enteric glia.

Enteric glia, the support cells of myenteric ganglia, have been widely studied with respect to their morphology and immunohistochemical phenotype, but little is known about their functional properties. We developed a method for the amplification of enteric glia from newborn guinea pigs to further characterize these cells. Treatment with a combination of basic fibroblast growth factor and the adenylate cyclase activator, cholera toxin, permitted expansion of enteric glial cultures to confluence and serial passage for up to 8 months. The long-term cultured cells retained expression of 1) S100 protein, 2) GD3 ganglioside recognized by the monoclonal antibody LB1, and 3) the gene encoding glutamine synthetase. The electrophysiologic properties of cultured enteric glia were studied under whole-cell patch clamp conditions. Most cells expressed "delayed rectifier"-type potassium currents, and some also demonstrated tetrodotoxin-sensitive sodium currents. Other subsets of voltage-dependent potassium currents, calcium currents, and glutamate-gated currents were not demonstrable.

Animals↗
PubMed1993

Early management of younger adults dying of community acquired pneumonia.

We identified all patients under the age of 65 dying from CAP over a 3-yr-period in hospital in our health district. Most had chronic underlying illnesses. The early management of those who had previously been in good health was studied in greater detail, and was not ideal. Recommendations have been made to try and improve the assessment and treatment of patients with severe community-acquired pneumonia.

Adult↗
PubMed1993

The isolation and structural elucidation of four novel triterpene lactones, pseudolarolides A, B, C, and D, from Pseudolarix kaempferi.

Four novel triterpene lactones, pseudolarolides A [1], B [2], C [3], and D [4], were isolated from the seeds of Pseudolarix kaempferi. Their structures and stereochemistry were elucidated from spectral data. Compound 2 shows potent cytotoxicity against three human cancer cell lines, KB (nasopharyngeal), A-549 (lung), and HCT-8 (colon), and against a murine leukemia cell line (P-388) with ED50 values of 0.49, 0.67, 0.73, and 0.79 micrograms/ml, respectively.

Animals↗
PubMed1993

The alkaline protease of Aspergillus fumigatus is not a virulence determinant in two murine models of invasive pulmonary aspergillosis.

Little is known of the pathophysiology of invasive pulmonary aspergillosis (IPA), an opportunistic fungal infection usually caused by Aspergillus fumigatus. It has been suggested that the ability of the fungus to degrade elastin may aid its invasion and growth in lung tissue. We have described previously the construction of a strain of A. fumigatus in which the gene encoding an alkaline protease, AFAlp, had been disrupted (C.M. Tang, J. Cohen, and D.W. Holden, Mol. Microbiol. 6:1663-1671, 1992); this mutant is deficient in extracellular proteolytic and elastinolytic activity over a broad pH range. In this study, we compared the pathogenicity of this and another AFAlp disruptant with their isogenic, elastase-producing parental strains in two murine models of IPA. In both models, animals were inoculated via the respiratory tract. In the first model, the inoculum was delivered as airborne conidia and animals developed signs of respiratory distress within 2 to 4 days. In the second model, conidia were administered intranasally as a suspension and the disease developed over a 2-week period. No difference was observed between the wild-type and AFAlp disruptants in terms of mortality, and elastin breakdown was detected in lung tissue from animals inoculated with all four strains. We conclude that AFAlp is not a virulence determinant in these models of IPA.

Animals↗
PubMed1993

The detection of Aspergillus spp. by the polymerase chain reaction and its evaluation in bronchoalveolar lavage fluid.

Invasive pulmonary aspergillosis (IPA) is an important cause of mortality and morbidity in the immunocompromised host. However, the diagnosis of this condition may be difficult, and it is sometimes missed because of the lack of sensitivity of available tests. Therefore, we used polymerase chain reaction (PCR)-based amplification of fragments of genes-encoding alkaline proteases from Aspergillus fumigatus and A. flavus to detect these organisms in bronchoalveolar lavage fluid specimens. The predicted size of the product (747 base pairs) after amplification of A. fumigatus was larger than that for A. flavus (690 base pairs). The reaction was highly sensitive (after amplification of 500 fg of A. fumigatus DNA, product could be detected by Southern analysis), and it was specific for A. fumigatus and A. flavus. Bronchoalveolar lavage fluid from four immunosuppressed patients with proved or probable IPA was positive by this assay (sensitivity, 100%); in addition, the sample from one patient with possible IPA was PCR-positive. Only one specimen from 18 immunosuppressed patients with no evidence of IPA was PCR-positive (specificity, 94.4%). Five of 28 bronchoalveolar lavage samples from nonimmunosuppressed patients were PCR-positive, probably representing colonization of the respiratory tract. PCR-based detection may prove useful in the diagnosis of IPA.

Adult↗