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D Han

Publications and source records attributed to D Han.

At least 127 records · Page 7Linked to original sources

Neurotransmitter modulation of gap junctional communication in the rat hippocampus.

Increasing experimental evidence indicates that gap junctions can be modulated by neurotransmitters, in particular dopamine. To examine possible modulation of gap junctional communication in the rat hippocampus by neurotransmitters, we studied dye coupling and electrotonic transmission in the CA1 area in the presence of carbachol, a cholinergic agonist, and dopamine agonists. Carbachol markedly reduced dye coupling and the frequency of electrotonic potentials (spikelets). Spikelet amplitudes were decreased in the presence of carbachol. These effects were reversed by the cholinergic antagonist atropine, suggesting a muscarinic action of carbachol on gap junctional function. The non-specific dopamine agonist apomorphine, and the specific D1 receptor agonist SKF 38393, reduced dye coupling between pyramidal cells. Spikelet frequency was also decreased in the presence of dopamine agonists, but less than with carbachol. The specific D1 receptor antagonist, SCH 23390, reversed the effects of both dopamine agonists. These observations indicate that cholinergic and dopaminergic transmission can affect electrical and chemical (dye coupling) communication through gap junctions, and could therefore alter properties of neuronal assemblies, in addition to their effects on intrinsic membrane properties.

2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-ben↗

A two year prospective study to compare culture and polymerase chain reaction amplification for the detection and diagnosis of Lyme borreliosis.

AIM: To compare polymerase chain reaction (PCR) amplification of borrelial DNA and culture isolation of spirochaetes for the diagnosis of Lyme borreliosis by direct detection of Borrelia burgdorferi sensu lato in patients with erythema migrans and acrodermatitis chronica atrophicans lesions. METHODS: Skin biopsy specimens from erythema migrans and acrodermatitis chronica atrophicans lesions were subdivided and tested by PCR amplification assay and culture using two artificial growth media, Barbour-Stoenner-Kelly II (BSK II) and modified Kelly-Pettenkofer (MKP). Five classes of lesions were studied: typical erythema migrans, spontaneously resolved erythema migrans, atypical/partially treated erythema migrans, typical acrodermatitis chronica atrophicans, and atypical/partially treated acrodermatitis chronica atrophicans. RESULTS: For both erythema migrans and acrodermatitis chronica atrophicans lesions, the most sensitive detection method was MKP culture. PCR was less sensitive than MKP culture, but more sensitive than BSK II culture. Results for 758 typical erythema migrans specimens showed positivity rates of 36% for MKP, 25% for PCR, and 24% for BSK II. Differences were statistically significant. The overall positivity rate for all three methods combined was 54%, but few specimens (6%) were positive by all three methods. Examination of multiple erythema migrans lesions from the same patient increased the diagnostic yield. These findings, and similar results for acrodermatitis chronica atrophicans lesions, suggest that the distribution of spirochaetes in skin biopsies is not homogeneous. CONCLUSIONS: Although possessing the potential to provide a rapid diagnosis, PCR is not more sensitive than culture for the direct detection of borrelia. Spirochaetes appear to be unevenly distributed throughout biopsy specimens, suggesting that diagnosis of Lyme borreliosis by direct detection of the causative agent in skin lesions in vulnerable to sample bias.

Acrodermatitis↗

Protection against glutamate-induced cytotoxicity in C6 glial cells by thiol antioxidants.

In many cell lines, glutamate cytotoxicity is known to be medicated by an inhibition of cystine transport. Because glutamate and cystine share the same transporter, elevated levels of extracellular glutamate competitively inhibit cystine transport leading to depletion of intracellular glutathione. A glutathione-depleted state impairs cellular antioxidant defenses resulting in oxidative stress. It was therefore of interest to investigate whether proglutathione agents, e.g., N-acetylcysteine and lipoic acid, are able to protect against glutamate cytotoxicity. Both lipoic acid (100 microM-1 mM) and N-acetylcysteine (100 microM-1 mM) completely protected C6 cells from the glutamate-induced cell death. Both agents facilitate extracellular supply of cysteine, the reduced form of cystine, that is transported into the cell by a glutamate-insensitive transport mechanism. Protection by lipoic acid and N-acetylcysteine corresponded with a sparing effect on cellular glutathione, which is usually depleted after glutamate treatment. In the presence of L-buthionine-(S,R)-sulfoximine, a gamma-glutamylcysteine synthetase inhibitor, low doses (< 100 microM) of lipoic acid and N-acetylcysteine did not protect cells against glutamate-induced cytotoxicity. At higher concentrations (> 500 microM), however, both lipoic acid and N-acetylcysteine provided partial protection against glutamate cytotoxicity even in glutathione synthesis-arrested cells. These results indicate that at low concentrations the primary mechanism of protection by the thiol antioxidants was mediated by their proglutathione property rather than direct scavenging of reactive oxygen. At higher concentrations (> 500 microM), a GSH-independent direct antioxidant effect of lipoic and N-acetylcysteine was observed. Dichlorofluorescin fluorescence, a measure of intracellular peroxides, increased sixfold after glutamate treatment of C6 cells. Lipoic acid and N-acetylcysteine treatment significantly lowered glutamate-induced dichlorofluorescin fluorescence compared with that of controls. Interestingly, alpha-tocopherol (50 microM) also suppressed glutamate-induced dichlorofluorescin fluorescence, indicating the peroxides detected by dichlorofluorescin were likely lipid hydroperoxides. Both thiol antioxidants, particularly lipoic acid, appear to have remarkable therapeutic potential in protecting against neurological injuries involving glutamate and oxidative stress.

Acetylcysteine↗

Effect of glutathione, catechin, and epicatechin on the survival of Drosophila melanogaster under paraquat treatment.

The biological effect of antioxidants which showed high superoxide-scavenging (SOS) activity in an in vitro analysis was examined by using Drosophila melanogaster. When the flies were exposed to paraquat as an endogenous source of the superoxide anion, their survival rapidly decreased. Although the SOS antioxidants did not have a preventive effect against paraquat toxicity, a supplement of each SOS antioxidants as glutathione, (+)-catechin and/or (-)-epicatechin to the diet had a reparative effect on flies damaged by the superoxide anion. The survival ratio of flies fed on a diet enriched with SOS antioxidants ranged from 77% to 87%, while that of the control group was 56%. When flies were exposed to paraquat in the presence of hydrogen peroxide or iron, each combination was more toxic than paraquat alone, since the two compounds could accelerate the generation of reactive oxygen species in vivo. The SOS antioxidants, however, allowed the flies to resist the combined toxicity of paraquat and ferrous iron.

Animals↗

Flat depressed early colon cancer--a case report.

A flat depressed early colon cancer (FDEC) is characterized by non-polypoid growth pattern, no association of adenomatous tissues and a tendency of even small lesions toward submucosal invasion and lymph node metastasis. It supports de novo carcinogenesis of colorectal cancer, although most colorectal cancers arise in pre-existing adenoma (adenoma-carcinoma sequence). There have been few reports of small depressed cancers because of the difficulty in colonoscopic detection and the rapid development to ulcerating advanced cancers. We report a case of flat depressed early colon cancer confined to mucosa detected by indigo carmine contrast colonoscopy.

Colonic Neoplasms↗

[Scanning electron microscopic observation of the three-dimensional structure of laryngeal carcinoma using freeze-cracking method].

OBJECTIVE: To investigate the relation between the morphology and the biological characteristics of laryngeal carcinoma. METHODS: The freeze-cracking method was used to examine 126 specimens of the larynx under scanning electron microscope (SEM). The specimens were obtained from 12 patients suffering from laryngeal cancer and 4 patients with normal larynx. RESULTS: (1) In the normal larynx, the squamous-cell epithelium showed marked keratinization and the intracellular bridges were clear and regular. (2) In areas adjacent to cancer, the structure of the squamous-cell epithlium could be identified, but the cells were deranged and paramorphic. (3) There were two types in the marginal area of the carcinoma. Along the areas adjacent to cancer, infiltrating lymphocytes were usually observed and the surface of the lymphocytes had many leading lamella. CONCLUSION: The results indicate that the three-dimensional observation with SEM using the freeze-cracking technique is helpful for studying the relation between morphology and biologic characteristics of laryngeal carcinoma.

Carcinoma, Squamous Cell↗

[The study of effects of immunologic modifier on proliferation and cytotoxicity against HEP-2 of LAK cells in vitro].

Adoptive immunotherapy as a new approach has been used to treat advanced carcinoma for almost ten years, and gained some good results. However, the application of this method is largely restricted by the side effects along with high dose of IL-2 and lack of sufficient amount of LAK cells. Mycobacterial Polysaccharides (MPS) and anti-CD3 antibodies (CD3Ab) have been demonstrated to be effective immunologic modifiers capable of proliferating lymphocytes and increasing cytotoxity against tumor cells. In this article we discussed the effects of MPS and CD3Ab on proliferation and cytotoxity against HEP-2 cells of LAK cells. Our results showed that: 1. MPS group (incubating IL-2 2 x 10(5)/L plus MPS 0.4 mg/L) or CD3 group (IL-2 1 x 10(6)/L plus CD3Ab 4 mg/L) were more proliferative than LAK group (only incubated with IL-2 1 x 10(6)/L). MPS group is the most proliferative in the three groups, P < 0.05. 2. Cytotoxity against HEP-2 in MPS or CD3 group was also better than in LAK group. CD3 group could maintain good cytotoxity in the longest period. These results could provide guidance for adoptive immunotherapy assisted by MPS or CD3Ab.

Antibodies, Monoclonal↗

[The observation on treatment effects of local adoptive immunotherapy in 33 cases with head and neck cancer].

OBJECTIVE: To evaluate treatment effects of local adoptive immunotherapy in 33 cases with head and neck cancer. METHODS: IL-2 1 x 10(5)-2 x 10(5) U per day was injected into tumor for 10 days; during the 4th-8th day LAK cells 1.0 x 10(8)-5.0 x 10(8) per day were combined with IL-2 injection. Then we analysized clinical effect, immunopathology, side effects, etc. RESULTS: CR was seen in 1 case; PR in 6, MR in 20 cases, and SD in 6 cases. The one-, two- and three-year survival rate was 96.3%, 83.3% or 75.0% respectively. The results of histopathology showed that there were large amount of T lymphocytes (CD3+/4+) infiltrating in the tumor area after immunotherapy. No serious side effects were found during the course of the treatment. CONCLUSION: Local injection of IL-2/LAK can be used as immunotherapy for head and neck cancer.

Adolescent↗

Mutation effect of high altitude balloon flight on rice and green pepper seeds.

Dry seeds of rice and green pepper were carried in the basket of a high altitude balloon to 30-40 km for 8 h. The treated seeds were planted on the ground and characters of these seeds and their progenies, such as development, fruiting, disease resistance were observed. The results showed that the SP2 generation of the treated rice showed strong mutation. A Japonica male fertility restorer rice line was found out. It could be hybridized with sterile line of Indica rice, and their progeny had strong hybrid vigor, high setting percentage and good plumpness of grain. Disease resistance, high-yield, big-fruit and high vitamin C content strains were selected from the progenies of treated green pepper seeds. The above results indicated that the special conditions in high altitude could produce a variety of mutations and most of them are hereditable. Therefore, high altitude balloon flight can be used as a new method of mutation breeding.

Agriculture↗

Temporal specificity of muscarinic synaptic modulation of the Ca(2+)-dependent K+ current (ISAHP) in rat hippocampal neurones.

1. We examined synaptic modulation of the Ca(2+)-dependent K+ current (ISAHP), which underlies the slow after-hyperpolarization (sAHP) in hippocampal CA1 neurones of rat brain slices. ISAHP was evoked in whole-cell voltage-clamp mode by depolarizing pulses, and synaptic afferents to CA1 neurones were stimulated electrically with a paired-pulse protocol. 2. Afferent stimulation delivered 200-1500 ms prior to be depolarizing pulse produced a profound reduction of ISAHP by 58%, but not other Ca(2+)-dependent outward currents that preceded ISAHP. Perfusion of slices with atropine significantly attenuated the synaptic reduction of ISAHP, indicating an event mediated largely by muscarinic receptor activation. When delivered < 400 ms after the depolarizing pulse, similar synaptic stimuli produced no substantial reduction in ISAHP, even in neurons where the duration of ISAHP was prolonged to 8-10 s either by lowering the recording temperature or by intracellular application of a calcium chelator. 3. To examine the effect of cholinergic stimulation of the depolarization-activated Ca2+ influx, high-threshold voltage-activated Ca2+ currents were recorded in the conventional or perforated whole-cell mode. Perfusion of slices with 5-10 microM carbachol for 5-10 min caused no substantial decrease in these Ca2+ currents, suggesting that the synaptic reduction of ISAHP is unlikely to be due to a blockade of depolarization-induced Ca2+ influx which triggers the generation of ISAHP. 4. The present data demonstrate that afferent stimulation reduces ISAHP only if it occurs prior to the depolarization-induced Ca2+ influx. We propose that modulation of inactive sAHP channels by muscarinic stimulation may decrease their sensitivity to the influx of Ca2+, whereas sAHP channels activated by Ca2+ may compete with the receptor-coupled modulation thus rendering the sAHP channels unresponsive to cholinergic afferent stimulation.

Animals↗

Evolution of proneural atonal expression during distinct regulatory phases in the developing Drosophila eye.

BACKGROUND: Receptors of the Notch family affect the determination of many cell types. In the Drosophila eye, Notch antagonises the basic helix-loop-helix (bHLH) protein atonal, which is required for R8 photoreceptor determination. Similar antagonism between Notch and proneural bHLH proteins regulates most neural cell determination, however, it is uncertain whether the mechanisms are similar in all cases. Here, we have analyzed the sensitivity of atonal expression to Notch signalling using a temperature-sensitive Notch allele, by the expression of activated Notch or of the ligand Serrate, and by monitoring expression of the atonal-dependant gene scabrous and of the Notch-dependent Enhancer of split genes. RESULTS: The atonal expression pattern evolves from general "prepattern' expression, through transient "intermediate groups' to R8 precursor-specific expression. Successive phases of atonal expression differ in sensitivity to Notch. Prepattern expression of atonal is not inhibited. Inhibition begins at the intermediate group stage, corresponding to the period when atonal gene function is required for its own expression. At the transition to R8 cell-specific expression, Notch is activated in all intermediate group cells except the R8 cell precursor. R8 cells remain sensitive to inhibition in columns 0 and 1, but become less sensitive thereafter; non-R8 cells do not require Notch activity to keep atonal expression inactive. Thus, Notch signaling is coupled to atonal repression for only part of the atonal expression pattern. Accordingly, the Enhancer-of-split m delta protein is expressed reciprocally to atonal at the intermediate group and early R8 stages, but is expressed in other patterns before and after. CONCLUSIONS: In eye development, inhibition by Notch activity is restricted to specific phases of proneural gene expression, beginning when prepattern decays and is replaced by autoregulation. We suggest that Notch signalling inhibits atonal autoregulation, but not expression by other mechanisms, and that a transition from prepattern to autoregulation is necessary for patterning neural cell determination. Distinct neural tissues might differ in their proneural prepatterns, but use Notch in a similar mechanism.

Animals↗

IL2 triggers a tumor progression process in a melanoma cell line MELP derived from a patient whose metastasis increased in size during IL2/INFalpha biotherapy.

Human melanomas may express both in vivo and in vitro functional IL-Rs and may be expected to directly respond to injected IL2. This may generate biological situations which may be favourable for the patient, but also for tumor progression. Here, we analyse the latter hypothesis. MELP is a melanoma cell line derived from a patient whose metastasis increased in size during IL2/IFN alpha biotherapy [correction of biotheraphy]. These cells have been characterized in vitro for their phenotype and for their sensitivity to IL2. In vitro MELP cells express an IL2-R alpha(+) beta(+) gamma(-) phenotype and IL2 treatment induces the acquisition of new functional characteristics represented (i) by the increased surface expression of two markers of metastatic evolution (ICAM-1 and CD44); (ii) by the stable induction of the IL2-R gamma with the appearance of functional IL2-R beta complex, which are also recognized by GM-CSF; (iii) by the inhibition of transcription of a regulatory cytokine such as IL6; (iv) by a differential effect of IL6 on CD44 surface expression in MELP cells treated or not with IL2 (MILG cells); (v) by the acquisition of faster growth rates and appearance of piling up and multilayer cellular organization; (vi) by the development of rapidly growing tumors in nude mice. IL2 induces in MELP cells a tumor progression process that could mimic the metastatic evolution observed in vivo during biotherapy. Therefore, MELP phenotype may help to define a subset of patients in which IL2 therapy may trigger unfavourable evolution.

Adult↗

Single-tube nested polymerase chain reaction assay based on Flagellin gene sequences for detection of Borrelia burgdorferi sensu lato.

An inherent drawback of nested PCR systems to increase sensitivity of PCR-based assays is that tubes must be opened after the first round of amplification in order to transfer template molecules to the second amplification reaction; this procedure introduces the risk of carry-over contamination of negative specimens. To obviate this disadvantage, a nested PCR assay for detection of Borrelia burgdorferi in which both amplifications are performed in a single tube that remains closed throughout the entire process was devised. The assay is based on flagellin gene sequences with previously determined species-wide and species-specific properties. The nested PCR system proved to be 1000 times more sensitive than the conventional assay. Using the nested PCR system, ten spirochaetes could be routinely detected by agarose gel electrophoresis alone, whereas the conventional PCR system could detect only 10(4) spirochaetes under these conditions. After Southern transfer on amplification products and hybridization with 320- or chemiluminescent-labeled probes, the nested PCR system could easily detect a single spirochaete by both means, whereas the sensitivity of the conventional PCR assay varied from 10(1) (32P) to 10(3) (chemiluminescence) spirochaetes. This single-tube nested PCR system should be a useful addition to the current range of diagnostic assays for Lyme borreliosis.

Base Sequence↗

Molecular characterization of Borrelia burgdorferi sensu lato from Slovenia revealing significant differences between tick and human isolates.

One hundred twenty-nine Slovenian isolates of Borrelia burgdorferi sensu lato derived from patients (69 strains) or Ixodes ricinus ticks (60 strains) were characterized. All of the strains were first- or second-passage isolates obtained in 1992 and 1993 from the same endemic region. The techniques used for the molecular analysis of strains included species-specific polymerase chain reaction (PCR) typing, and pulsed-field gel electrophoretic separation of undigested and MluI-digested genomic DNA. Isolates were identified to the species level by large restriction fragment pattern (LRFP) analysis and the results compared with the species-specific PCR result. Fifty-two patient isolates (75%) were typed as Borrelia afzelii (LRFP MLa1), 6 (9%) as Borrelia garinii (LRFPs MLg1-4), and 11 (16%) as Borrelia burgdorferi sensu stricto. The latter included 9 isolates (13%) with a new LRFP that is not typical of Borrelia burgdorferi sensu stricto and for which the designation MLx is suggested. In contrast, only 32 of 60 (53%) tick isolates were typed as Borrelia afzelii, while 20 strains (33%) were typed as Borrelia garinii and 8 strains (13%) as Borrelia burgdorferi sensu stricto. Three new LRFPs were found among the Borrelia garinii (MLg5 and 6) and Borrelia burgdorferi sensu stricto (MLb15) tick isolates. Large restriction fragment pattern analysis identified new groups of Borrelia burgdorferi sensu lato and revealed an apparent difference in the isolation frequency of different species from patients and ticks in the same endemic region.

Animals↗

MR findings in human fascioliasis.

Fasciola hepatica is a trematode of herbivorous mammals, and man is infected by accidental ingestion of contaminated raw aquatic vegetables or water. There have been many reports on computed tomographic and cholangiographic features of human fascioliasis. However, findings of magnetic resonance (MR) imaging have not been reported. Hepatic fascioliasis produces three types of lesions in MR images arranged in tract-like fashion. The outermost area presents as an iso-signal area in T1WI, with slightly higher signal intensity in T2WI and diffuse enhancement after i.v. contrast. The second type presents as a well defined low signal area in T1WI, not enhanced, and also shows low signal intensity in T2WI. The third type has low signal intensity in T1WI, is not enhanced, and has high signal intensity in T2WI which is similar to fluid-containing inflammatory lesions such as pyogenic abscess. These findings suggest various changes associated with traumatic hepatitis caused by the migration of the worm in the liver. This diverse signal intensity can be a suggestive finding of fascioliasis.

Fascioliasis↗