Biomedical subjects

H Wang

Publications and source records attributed to H Wang.

At least 433 records · Page 24Linked to original sources

PubMed2001

The combination of stricture dilation, endoscopic needle aspiration, and biliary brushings significantly improves diagnostic yield from malignant bile duct strictures.

BACKGROUND: Brush cytology, routinely performed at ERCP to assess malignant-appearing biliary strictures, is limited by relatively low sensitivity and negative predictive value. This study assessed whether the combination of stricture dilation, endoscopic needle aspiration, and biliary brushing improves diagnostic yield. METHODS: In a prospective nonrandomized study, 46 consecutive patients were evaluated with malignant-appearing biliary strictures at ERCP. Twenty-four patients (Group A) underwent standard brush cytology alone and 22 patients (Group B) underwent stricture dilatation to 10F, endoscopic needle aspiration, and subsequent biliary brushing by using the Howell biliary system. The diagnostic yields for both techniques were compared. RESULTS: Of the 46 patients, 34 had proven malignant strictures (14 Group A, 20 Group B). Compared with brushing alone, the combination of stricture dilatation, endoscopic needle aspiration, and subsequent biliary brushing significantly increased both the sensitivity (57% vs. 85%, p < 0.02) and specificity (80% vs. 100%, p < 0.02) of cytology with positive brushings in all patients with pancreatic or gallbladder carcinoma. CONCLUSIONS: The combination of stricture dilation, endoscopic needle aspiration, and biliary brushing significantly improves diagnostic yield for malignant bile duct strictures and may particularly be of benefit for extrinsic strictures caused by pancreatic or gallbladder carcinoma.

Adult↗

PubMed2001

Cigarette smoke inhibits osteogenic differentiation and proliferation of human osteoprogenitor cells in monolayer and three-dimensional collagen gel culture.

Cigarette smoke is a risk factor not only for emphysema but also for other disorders characterized by deficient tissue repair, including osteoporosis. We hypothesized, therefore, that smoke might directly impair bone cell repair processes. To evaluate this, bone marrow osteoprogenitor cells were isolated from normal subjects and cultured in monolayer and in three-dimensional type I collagen gel culture. Human osteoprogenitor cells could be induced to differentiate toward osteoblast-like cells in both culture conditions by osteogenic supplements. Under both culture conditions, cigarette smoke extract (CSE) inhibited the proliferation of osteoprogenitor cells in a concentration-dependent manner. CSE also inhibited differentiation of osteoprogenitor cells toward osteoblast-like cells as assayed by alkaline phosphatase activity and calcium incorporation into cell layer. Cells in monolayer culture were more sensitive to the effect of smoke than cells in three-dimensional gel culture. Similar results were obtained with osteoblast-like cells derived from osteosarcomas. This study, therefore, demonstrates that cigarette smoke may affect bone progenitor cells directly and in this manner may contribute to the development of osteoporosis.

Cell Differentiation↗

PubMed2001

The in vivo effect of red clover diet on ventral prostate growth in adult male mice.

The incidence of clinically significant prostate disease in Asian men is lower than in Western men and appears to be influenced by dietary and environmental factors. The aim of this study was to examine the effect of dietary isoflavones on prostate growth in intact male mice using an extract of red clover. The results demonstrated that prostate, but not testis, size was significantly reduced over 28 days of a red clover isoflavone supplemented diet. Histological examination revealed an increase in apoptotic cells, rather than a reduction in proliferative activity in the epithelium. These findings support the hypothesis that dietary red clover isoflavones can induce apoptosis and lead to a reduction in prostate size.

Animal Feed↗

PubMed2001

RPA facilitates rejoining of DNA double-strand breaks in an in vitro assay utilizing genomic DNA as substrate.

PURPOSE: Replication protein-A (RPA) is a heterotrimeric single-stranded DNA-binding protein playing essential roles in many aspects of nucleic acid metabolism, including DNA replication, nucleotide excision repair and homologous recombination. Here, the role of RPA in the rejoining of radiation-induced DNA double-strand breaks (DSB) by non-homologous end-joining (NHEJ) was investigated. METHODS AND MATERIALS: A previously described in vitro assay for DSB rejoining was employed. The assay used 'naked' genomic DNA prepared from agarose-embedded G(1)-phase A549 cells as a substrate and extracts prepared from HeLa cells as a source of enzymes. Rejoining of DSB in this assay is absolutely dependent on cell extract and proceeds, under optimal reaction conditions, to an extent similar to that observed in intact cells. For experiments, extracts were supplemented with excess purified recombinant RPA. Alternatively, RPA was removed from the extracts either by fractionation or immunodepletion. RESULTS: Although the rejoining of DSB in vitro was not absolutely dependent on RPA, it proceeded faster and to higher levels of completion when recombinant protein was added to the extracts. Depletion of RPA from extracts reduced the rejoining half-times and addition of purified recombinant protein restored the kinetics of DSB rejoining. Extract fractionation indicated the operation of at least two pathways in DSB rejoining, only one of which was facilitated by RPA. CONCLUSIONS: The results suggest that in addition to its role in homologous recombination, RPA may also have a supportive role in some forms of non-homologous end-joining.

DNA↗

PubMed2001

Remedial strategies for municipal solid waste management in China.

The purpose of this investigation is to evaluate the current status and to identify the problems of municipal solid waste (MSW) management in China to determine appropriate remedial strategies. This is the second of two papers proposed on this topic. Major problems or difficulties identified in MSW management in China include MSW land, air, and water pollution, commingled collection, poor administration, shortage of funds, lack of facilities, and problems of training and public awareness. In order to solve these problems and to improve MSW management in China, remedial strategies in three areas are recommended: institutional reform, technology development, and legislation and administrative improvement. The primary principle involved in institutional reform is unifying legislative responsibilities into one body and developing a market mechanism for handling MSW. Composting, landfills, and incineration should be equally developed in accordance with China's needs. The feasibility of developing technology to handle MSW in China is discussed. Also recommended is the establishment of sound regulatory systems, including a service fee system, a source separation system, and a training program. China is presently undergoing economic and institutional reform at the national and local levels. Results of this study will provide useful information on MSW management in China.

China↗

PubMed2001

Municipal solid waste characteristics and management in China.

The purpose of this investigation is to evaluate the current status and identify the problems of municipal solid waste (MSW) management in China in order to determine appropriate remedial strategies. This is the first of two papers, discussing primarily the general characteristics of MSW and its management in China. The second paper focuses on specific remedial strategies. MSW generation in China has increased rapidly in the past 20 years from 31.3 million tons in 1980 to 113.0 million tons in 1998. The annual rate of increase is 3-10%. The average generation per capita is 1.0 kg/day (0.38 t/year). Nearly one-half of the waste generated is dumped in the suburbs, where the accumulated quantity has reached 6 billion tons, which has caused heavy environmental pollution. This paper provides information on MSW management in China, such as MSW generation and its physical, chemical, and biological properties. Low calorific value and high moisture content characterize China's municipal waste. Other issues related to MSW management in China are also discussed, including the factors that influence MSW generation quantity and

China↗

PubMed2001

Prevalence of serotypes and molecular epidemiology of Streptococcus pneumoniae strains isolated from children in Beijing, China: identification of two novel multiply-resistant clones.

Three-hundred and seventy-six strains of Streptococcus pneumoniae isolated from clinical specimens and nasopharyngeal swabs from children at daycare centers and hospitals in Beijing China, between January 1997 and March 1998, were serotyped. Twenty-seven different serotypes were identified. The most prevalent serotypes in the carriage isolates were 6A, 19F, 23F, and 15 and were found in 66.8% of cases. Serotype data indicate that 51.8% of carrier strains would be included in the 11-valent conjugate vaccine formulation, while inclusion of vaccine-related serotypes, increased the potential vaccine coverage to 79.4%. Serotypes 7, 6B, 23F, 19F, 15, and 3 accounted for 62% of clinical strains, with 70% vaccine-related serotypes. DNA fingerprinting of 47 penicillin resistant and 71 penicillin-susceptible/macrolide-resistant strains by BOX polymerase chain reaction (PCR), pulsed-field gel electrophoresis (PFGE), and penicillin binding protein (PBP)-fingerprinting identified two novel clones: one a serotype 23F multiresistant clone resistant to penicillin, tetracycline, erythromycin, clindamycin, and variably resistant to chloramphenicol and trimethoprim-sulphamethoxazole; and the second a multiresistant penicillin-susceptible, macrolide-resistant serotype 6A clone, highly resistant also to tetracycline, clindamycin, and trimethoprim-sulphamethoxazole. The macrolide resistance determinant in 89% of erythromycin-resistant strains tested (penicillin-susceptible and penicillin-resistant) was the erm gene, both the erm and mef genes were simultaneously found in 6%, and mef alone in 3.4%. The data demonstrates that macrolide resistant strains in China include clonal strains and strains with dual mef and erm resistance determinants.

Anti-Bacterial Agents↗

PubMed2001

N-hydroxysuccinimidyl-fluorescein-O-acetate for precolumn fluorescence derivatization of amino acids and oligopeptides in liquid chromatography.

A new fluorescent derivatizing reagent, N-hydroxysuccinimidyl-fluorescein-O-acetate, is used for the high-performance liquid chromatographic analysis of amino acids and oligopeptides. This reagent has the advantages of high-detection sensitivity in the visible region, specifically with amino groups, mild derivatization conditions, and little interference induced. The fluorescence properties of the reagent and its derivatives with amino acids and oligopeptides are studied. The conditions of the derivatization are investigated in detail. In the mobile phase of methanol-water (42:58, v/v) containing a 10 mM pH 5.0 citric acid-Na2HPO4 buffer, six amino acids and oligopeptides are separated in 20 min with fluorescence detection at excitation and emission wavelengths of 492 and 513 nm, respectively, with the detection limits for injected standards ranging from 0.64 to 12 fmol.

Amino Acids↗

PubMed2001

Maturation of rhesus monkey oocytes in chemically defined culture media and their functional assessment by IVF and embryo development.

This study compared success of in-vitro maturation of rhesus monkey oocytes in protein-free versus serum-containing culture systems, assessed by embryo development subsequent to IVF. Four media were tested: (i) modified Connaught Medical Research Laboratories medium (mCMRL-1066); (ii) hamster embryo culture medium-10 (HECM-10); (iii) control: mCMRL-1066 + 20% bovine calf serum (BCS); (iv) HECM-10 + 20% BCS. Immature oocytes from FSH-stimulated rhesus monkeys were allocated among the media containing ovine FSH (5 microg/ml) and LH (10 microg/ml) and cultured for 36-40 h. Metaphase II ova were inseminated and putative zygotes were cultured in mCMRL-1066 + 20% BCS until development arrested. Ova matured in all four media had similar (P> 0.05) potential to initiate (66, 67, 82 and 69% respectively) and complete meiotic maturation (60, 50, 76 and 57% respectively). Inseminated ova in all groups had similar potential to be fertilized (86, 83, 84 and 90% respectively), cleave (71, 83, 76 and 90% respectively) and develop to the blastocyst stage (19, 17, 16 and 30% respectively). These results indicate for the first time that primate oocytes can be successfully matured in protein-free medium, with subsequent blastocyst development, comparable to responses in complex medium with serum. This finding will facilitate studies on mechanisms regulating primate oocyte maturation.

Animals↗

PubMed2001

CpG oligodeoxynucleotide vaccination suppresses IgE induction but may fail to down-regulate ongoing IgE responses in mice.

Antigen-specific IgE plays an important role in the pathogenesis of allergic disorders. Immunostimulatory CpG motifs (CpG) in bacterial DNA or synthesized oligodeoxynucleotides (ODN) are gaining recognition as potential immunomodulators for switching on protectiveT(h)1-mediated immunity and preventing or potentially inhibiting T(h)2-dependent allergic responses. To date, allergic models used in CpG ODN studies have been established by immunization of mice with allergen in the presence of adjuvant. This, in addition to failure to assess specific IgE production in most of the studies, has limited understanding of the role of CpG ODN vaccination in allergic responses. Here, we examine the effects of synthesized CpG ODN on both developing and ongoing IgE responses in mice sensitized using a recombinant mosquito salivary antigen (rAed a 2) without adjuvant. Pretreatment of mice with CpG ODN mixed with rAed a 2 successfully inhibited subsequent induction of serum rAed a 2-specific IgE (but not IgG1) and antigen-induced IL-4 and IL-5 production in spleen cells. This was associated with an increase of serum IgG2a and IL-12, and increased IFN-gamma and IL-12 production by spleen cells. In this model, however, co-administration of CpG ODN with rAed a 2 to presensitized mice failed to down-regulate ongoing IgE responses despite significant up-regulation of serum IL-12 and specific IgG2a. Strikingly, a transient skin delayed-type hypersensitivity reaction occurred in CpG ODN-treated mice. These observations provide a new insight into the potential therapeutic application of CpG ODN to allergic disorders.

Adjuvants, Immunologic↗

PubMed2001

Different regulation of oestrogen receptors alpha and beta in the human cervix at term pregnancy.

During pregnancy, a cervical connective tissue remodelling takes place, clinically recognized as softening, effacement and dilatation. The role of oestrogens and their receptors (ER) in this process is not clear. ERalpha is a ligand-activated transcription factor involved in many physiological processes. The identification of a second oestrogen receptor, ERbeta, has led to a re-evaluation of oestrogen signalling and physiology. The aim of this study was to monitor the expression of the two ERs in the cervix from women at term pregnancy (TP) and after parturition (PP) compared with that of non-pregnant (NP). A solution hybridization assay showed that the level of ERalpha mRNA was significantly decreased in the PP group, when compared with the NP and TP groups. In contrast the ERbeta mRNA level was increased in the TP group compared with the NP and PP groups. These results were supported by reverse transcription-polymerase chain reaction (RT-PCR). Similar results were observed for the protein with immunohistochemistry. Intense ERbeta immunostaining was observed in neutrophils and the endothelial cells of blood vessels. In conclusion, this study supports a concept according to which oestrogen might be involved in the final remodelling of the cervix via the modulating effects of the two ERs. Furthermore, oestrogen may mediate some effects on cervical ripening via ERbeta present in the invading neutrophils. Further studies are needed to elucidate this finding.

Adult↗

PubMed2001

Co-localization of oestrogen receptor beta and leukocyte markers in the human cervix.

Cervical ripening during parturition is associated with rapid production of catabolic enzymes by invading leukocytes and increased collagen metabolism. The recruitment of leukocytes is regulated by various factors including inflammatory mediators, prostaglandins and matrix metalloproteinases. Sex steroids may be indirectly or directly involved in this process. This study aimed to evaluate the expression of oestrogen receptor beta (ER beta) in blood cells infiltrating the cervix during pregnancy and parturition. Cervical biopsies were obtained from term pregnant, post-partal and non-pregnant women. The ER beta protein and leukocyte markers CD45 and CD68 were evaluated by single and double labelling immunohistochemistry. Quantitative values were assessed using a microscope and a high-resolution camera connected to a computer with image analysis program. The number of CD45(+) and CD68(+) cells in the cervix increased in term pregnancy and post-partum compared with the non-pregnant state. The ER beta antigen was co-localized with CD45 leukocyte common antigen and CD68 macrophage specific antigen in blood leukocytes infiltrating the cervical tissue. The presence of ER beta in the cervical leukocytes suggests that oestrogen may directly regulate leukocyte functions in the cervix.

Antibodies, Monoclonal↗

PubMed2001

Effect of age and breeding season on the developmental capacity of oocytes from unstimulated and follicle-stimulating hormone-stimulated rhesus monkeys.

Effects of age and season on the developmental capacity of oocytes from unstimulated and FSH-stimulated rhesus monkeys were examined. Immature cumulus-oocyte complexes were matured in vitro in modified CMRL-1066 medium containing 20% bovine calf serum and subjected to in vitro fertilization followed by embryo culture. After fertilization, ova from unstimulated prepubertal monkeys displayed lower development to morula (4%) than those from unstimulated adult females (18% in breeding season and 22% in nonbreeding season). No developmental difference was found between ova from unstimulated adult monkeys in breeding and nonbreeding seasons. However, ova from FSH-primed prepubertal monkeys displayed greater development to blastocyst stage (54%) than those from adult monkeys in the breeding season (16%) and nonbreeding season (0%); and ova from FSH-primed adult females in the breeding season had significantly (P < 0.05) greater developmental competence than those obtained in the nonbreeding season (> or = morula stage, 54% vs. 3%; blastocyst stage, 16% vs. 0%). These data indicate that 1) rhesus monkey oocytes acquire developmental competence in a donor age-dependent manner, and 2) animal age and breeding season modulate the effect of FSH on oocyte developmental competence in the rhesus monkey.

Aging↗

PubMed2001

Expression of matrix metalloproteinase-2, -9, -14, and tissue inhibitors of metalloproteinase-1, -2, -3 in the endometrium and placenta of rhesus monkey (Macaca mulatta) during early pregnancy.

The extracellular matrix proteolytic machinery has long been recognized as one of the most important mechanisms for regulating trophoblast invasion. Matrix metalloproteinases (MMPs) are a group of proteases involved in this process, and their activities are regulated by tissue inhibitors of MMPs (TIMPs). In this study, we collected rhesus monkey uteri on Days 12, 18, and 26 of pregnancy and examined the mRNA expression of MMP-2, -9, -14, and TIMP-1, -2, -3, as well as the activities of MMP-2 and -9 by using in situ hybridization and gelatin zymography, respectively. The results showed that MMP-2 and -9 were expressed earlier than MMP-14 and TIMPs in the pregnant endometrium. MMP-14 and TIMP-2 mRNAs appeared in perivascular decidual cells earlier than MMP-2 mRNA. On Day 26 of pregnancy, placental villi expressed little MMP-2, -14, and TIMP transcripts but abundant MMP-9 mRNA. Furthermore, MMP-2, -9, -14, and TIMP-1, -2, -3 were highly expressed on the fetal-maternal border but were absent in the myometrium. TIMP-3 mRNA in the endometrium was specifically localized to some cells lining the outer membrane of several groups of arterioles. Combined with the results obtained by gelatin zymography, we found that active MMP-2 existed in the endometrium throughout these three phases, while MMP-9 showed considerable activities only on Days 18 and 26 of pregnancy. The data suggest key roles for MMP-2 and -9 in invasion of trophoblast cells into the endometrium and the development of the placenta and might indicate that these processes are regulated by MMP-14 and TIMP-1, -2, and -3.

Animals↗

PubMed2001

Neuronal nitric oxide synthase (NOS) regulates the expression of inducible NOS in rat small intestine via modulation of nuclear factor kappa B.

We previously reported that neuronal nitric oxide synthase (nNOS) is the predominant NOS in the intestine. Inducible NOS (iNOS), an enzyme involved in the inflammatory response, is regulated by cytokines via the transcriptional factor NF-kappaB. We examined a new mechanism of intestinal iNOS regulation with respect to the role of nNOS and its effect on NF-kappaB. Young Sprague-Dawley rats were treated for 4 days with 1) saline, 2) 7-nitroindazole (7-NI, specific nNOS inhibitor), 3) 7-NI + pyrrolidine dithiocarbamate (PDTC, NF-kappaB inhibitor), or 4) PDTC. Intestinal iNOS mRNA, NF-kappaB activity, and the tissue content of the regulatory IkappaBalpha were examined. We found that 7-NI-treated animals had higher intestinal NF-kappaB (p50-p65) activity, lower IkappaBalpha content, and increased intestinal iNOS mRNA, iNOS protein, and iNOS activity compared with controls. All of these changes were abolished when PDTC was given together with 7-NI. PDTC alone had no effect. 7-NI induces a delayed increase in intestinal myeloperoxidase activity (after elevation in NF-kappaB and iNOS), which could be abrogated by PDTC. We conclude that in normal rat small intestine, nNOS suppresses the gene expression of iNOS through NF-kappaB down-regulation and that nNOS suppression leads to IkappaBalpha degradation, NF-kappaB activation, and iNOS expression.

Animals↗

PubMed2001

Multivariate prognostic analysis of stage I(E) primary non-Hodgkin's lymphomas of the nasal cavity.

From January 1968 to December 1997, a total of 71 patients with stage I(E) (Ann Arbor staging system, 1971) primary non-Hodgkin's lymphomas of the nasal cavity received treatment in the Cancer Center of Sun Yat-Sen University of Medical Sciences. Thirty-seven lesions were limited to the nasal cavity (limited I(E)), whereas the other 34 were extended to the structure out of the nasal cavity (extended I(E)). Forty-four patients were treated with radiochemotherapy and 27 with radiotherapy alone. Kaplan-Meier methods were used in the survival analysis. Multivariate analysis was carried out using the Cox proportional hazard model. The 5- and 10-year survival rates were 71.85% and 59.67% for the patients with a complete response to irradiation, and both were 13.89% for the patients with residue lesions (p = 0.0004). The 5- and 10-year survival rates were 69.81% and 56.72% for limited I(E), and 40.65% and 35.57% for extended I(E) (p = 0.0047). The prognosis was better for those younger than 44 years (p = 0.0003). The 10-year survival rates for radiotherapy alone and combined radiochemotherapy are 52% and 75% for limited I(E) versus 37.58% and 45% for extended I(E) (p = 0.0644). B symptoms did not significantly affect clinical outcome (p = 0.729). Multivariate analysis showed that complete response of local lesion after radiotherapy, invasion of the primary tumor to adjacent structures, and patients' age were independent prognostic factors. Our study showed that radiotherapy is the main treatment method for the primary non-Hodgkin's lymphomas of the nasal cavity; the addition of chemotherapy may improve long-term survival. The local tumor response to radiotherapy, whether the extranasal structures were invaded, and patients' age were independent prognostic factors.

Adolescent↗

PubMed2001

Phenotype of Barrett's esophagus and intestinal metaplasia of the distal esophagus and gastroesophageal junction: an immunohistochemical study of cytokeratins 7 and 20, Das-1 and 45 MI.

The pathogenesis of short segment Barrett's esophagus (SSBE) and intestinal metaplasia (IM) of the gastroesophageal junction (IMGEJ) are poorly understood. Also, these conditions are difficult to distinguish from one another based solely on endoscopic and pathologic criteria. Therefore, the aim of this study was to evaluate the immunophenotypic features of SSBE and IMGEJ and to compare the results with lesions of known etiologies: long segment BE (LSBE) caused by reflux disease and Helicobacter pylori-induced IM of the gastric antrum (IMGA). Routinely processed mucosal biopsy specimens from 11 patients with LSBE, 17 with SSBE, 10 with IMGEJ, 16 with IMGA, 17 with a normal nonmetaplastic GEJ, and 7 patients with a normal gastric antrum were immunohistochemically stained with monoclonal antibodies to: Das1, an antibody shown to react specifically with colonic goblet cells; 45M1, an antibody that recognizes the M1 gastric mucin antigen; and cytokeratin (CK) 7 and 20, antibodies that have previously been reported to show specific staining patterns in BE versus IMGA. Also evaluated was nonintestinalized mucinous epithelium from LSBE, SSBE, and also the normal GEJ and gastric antrum. LSBE, SSBE, and IMGEJ showed similar prevalences of Das1 (91% versus 88% versus 100%) and 45M1 reactivity (100% versus 100% versus 100%), and a similar pattern of CK7/20 reactivity (diffuse strong CK7 staining of the surface and crypt epithelium, and strong surface and superficial crypt CK20 staining) (91% versus 94% versus 90%). In contrast, although 45M1 reactivity in IMGA (93%) was similar to that of the other three groups, IMGA showed a significantly lower prevalence of Das positivity (13%, p < 0.001), and only a 14% prevalence of the CK7/20 staining pattern that was predominant in the other three groups (p < 0.001). Das1, 45M1, and CK7/20 staining were similar in nonintestinalized "cardia-type" mucinous epithelium from LSBE, SSBE, and the GEJ, but all were distinct from the normal gastric antrum. In summary, the immunophenotypic features of SSBE and IMGEJ are similar and closely resemble those seen in classic LSBE, but are distinct from IMGA. This may indicate that IM in LSBE, SSBE and at the GEJ have similar biologic properties. Based on our data, SSBE and IMGEJ cannot be distinguished on the basis of their immunophenotype.

Antibodies↗

PubMed2001

Modulation by nucleotides of binding sites for [3H]glibenclamide in rat aorta and cardiac ventricular membranes.

Radioligand binding techniques were employed to determine the modulation by nucleotides of the specific [3H]glibenclamide (Gli) binding to rat aortic and cardiac ventricular preparations. Saturation analysis revealed a single binding site with K(D) value of 31.3 nM and Bmax of 180 fmol/mg wet weight in aortic preparations. We also observed that [3H]Gli bound reversibly and specifically to cardiac membranes. Unlabeled glibenclamide displaced [3H]Gli-specific binding of cardiac membranes completely with K(I) of 54.4 nM. In cardiac membranes, adenosine triphosphate (ATP), adenosine diphosphate (ADP), and uridine diphosphate (UDP) (from 0.01-5 mM) concentration dependently inhibited [3H]Gli binding independent of Mg2+. The values of K(I) were 0.47, 0.22, and 0.58 mM, respectively. However, in aortic preparations, [3H]Gli-specific binding was increased by ATP of 5 and 10 mM and showed a biphasic response to ADP. At concentrations to 1 mM, ADP inhibited binding; above 5 mM, the specific [3H]Gli binding was increased. UDP did not alter the binding up to 5 mM. In the presence of Mg2+ (20 mM), the inhibitory effects of ATP (0.01-1 mM) or ADP (0.01-5 mM) on the binding in cardiac membranes were abolished, whereas the facilitatory effects of ATP or ADP in aortic preparations were strengthened. Analysis of kinetics showed that the time of [3H]Gli association and dissociation in cardiac and aortic preparations was monophasic. The association was delayed with dissociation unchanged by ATP, ADP, and UDP of 1 mM, respectively, in cardiac membranes. In aorta, however, at the same concentration ATP accelerated association and retarded dissociation and vice versa for ADP. Association and dissociation were not changed by UDP of 5 mM. We conclude that ATP, ADP, and UDP are all major allosteric modulators of K(ATP) channels and they affect the antagonist binding to heart (sulfonylurea receptor 2A) and aorta (sulfonylurea receptor 2B) differently.

Adenosine Diphosphate↗