Rejection of cardiac xenografts by NK cells and macrophages.
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Biomedical subjects
Publications and source records attributed to Y Lin.
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HK31 (S565-K595) has previously been shown to encompass the binding domain for plasma prekallikrein (PK) within domain 6 of high molecular weight kininogen (HK). The complementary binding domain for HK within PK is mapped to PK56 (F56-G86), in the Apple 1 domain and to PK266 (K266-C295) in the Apple 4 domain. Isothermal titration calorimetry demonstrated that either PK peptide binds to HK31 in 1:1 stoichiometry. Binding of the alternate PK peptide into a ternary complex is facilitated nearly 2-fold. Fluorescence emission spectroscopy revealed that only the binding of PK56 caused a limited decrease in intrinsic tryptophane fluorescence emission intensity of HK31. We conclude that the two PK peptides bind to the HK peptide at different sites. To map the minimal sequence within HK31, truncated new peptides were tested for their ability to compete with HK for binding PK in a cell-free system. D567-T591, a 25-residue peptide which contains sufficient structural information for binding kallikrein in solution, blocked the binding of kallikrein to HK bound to endothelial cells and inhibited PK activation to kallikrein and the generation of kallikrein-activated urokinase on endothelial cell surfaces. HK-derived peptides could modulate excessive fibrinolysis and hypotension in sepsis and multiple trauma.
A rapid assay for the quantification of hepatitis B virus DNA in human serum was developed. The principle of the method combines competitive polymerase chain reaction (cPCR) (for the controlled amplification of hepatitis B virus DNA) and scintillation proximity assay (SPA) technology (for rapid detection and quantitation of PCR products). It also incorporates a reproducible and simple method for the preparation of serum DNA suitable for PCR amplification. The assay has a better linear dynamic range than traditional methods that use 32P to detect PCR products. It was applied to a range of hepatitis B virus (HBV) surface antigen positive (HBsAg + ) sera, and shown to be more sensitive than a commercially available HBV DNA kit.
Prenyl modification of proteins by farnesyl and geranylgeranyl isoprenoids occurs in a variety of eukaryotic cells. Culturing of Trypanosoma brucei in the presence of [3H]mevalonolactone (which is hydrolyzed in cells to give mevalonic acid, the precursor of protein prenyl groups) and an inhibitor of mevalonic acid biosynthesis leads to the radiolabeling of a specific set of proteins when analyzed by gel electrophoresis. T. brucei proteins were also labeled when cells were cultured in the presence of [3H]farnesol or [3H]geranylgeraniol, and each prenol labels a distinct set of proteins. Unlike mammalian cells, only a few T. brucei proteins of molecular weights similar to those of the mammalian Ras superfamily of GTPase (20-30 kDa) were labeled with [3H]farnesol or [3H]geranylgeraniol. When the 0-55% ammonium sulfate fraction of T. brucei cytosol was fractionated on anion exchange chromatography, protein farnesyltransferase (PFT) and protein geranylgeranyltransferase-I (PGGT-I) activities were detected and elute as two distinct peaks. Partially purified T. brucei PFT and PGGT-I display partly different specificities toward prenyl acceptor substrates from those of mammalian protein prenyltransferases. As shown previously, rat PFT utilizes proteins ending in CVLS and CVIM as efficient prenyl acceptors and rat PGGT-I utilizes proteins ending in CVLL and CVIM in vitro. On the contrary, T. brucei PFT farnesylates a protein ending in CVIM but not CVLS or CVLL, and T. brucei PGGT-I preferentially geranylgeranylates a protein ending in CVLL.
To estimate an annual number of patients treated for Moyamoya disease in Japan and to describe the clinico-epidemiological features, a nationwide epidemiological survey was conducted in 1995. The study consisted of two questionnaires, which were distributed to departments randomly selected, of neurosurgery, neurology and pediatrics in hospitals throughout Japan. The first questionnaire inquired the number of the patients treated in 1994 and the second one detailed clinico-epidemiological information of each patient reported. Following major epidemiological findings emerged from the study: (a) The total annual number of patients treated for Moyamoya disease was estimated as 3900 (95% confidence interval (CI) 3500-4400) in Japan 1994, with the prevalence and incidence rates of 3.16 and 0.35 per 100,000 population, respectively; (b) the sex ratio (females to males) of the patients was 1.8; (c) the peak of age distribution of the patients was observed in 10-14 years old and a smaller peak in their forties; (d) the age at onset was under 10 years old in 47.8% of the patients, but some had developed the disease at the age of 25-49 years; (e) family history of Moyamoya disease was found in 10.0% of the patients; and (f) about 75% of the patients had normal activity of daily life or working ability even before treatment. The present findings were quite comparable with those obtained in the previous nationwide epidemiological survey in 1990.
Hematopoietic neoplasms involving the breast, although less common than breast carcinoma, are often clinically indistinguishable from other breast tumors. Microscopically, these tumors can mimic primary carcinoma of the breast, especially in limited material such as needle biopsy specimens. Forty-five hematopoietic breast neoplasms including 21 primary non-Hodgkin's lymphomas (NHLs), 19 secondary NHLs, 1 undetermined NHL, 1 lesion secondary to Hodgkin's disease, and 3 granulocytic sarcomas were reviewed with regard to histologic subtype, morphologic features, and immunophenotype. The median age of patients at presentation was 67 years for those with primary NHLs and 61 years for those with secondary NHLs. The majority of lymphomas were intermediate or high grade. Diffuse large cell type was by far the most common histologic subtype. No lymphomas resembling lymphomas of mucosa-associated lymphoid tissue were identified in this series, suggesting that such lymphomas are rare in breast compared with other sites such as the gastrointestinal tract and lung. Four primary NHLs, 2 secondary NHLs, and 1 granulocytic sarcoma were initially misdiagnosed as carcinomas; three patients underwent radical mastectomy, and at least three other patients nearly received surgical treatment. One primary anaplastic large cell lymphoma of B-cell origin was identified that closely resembled poorly differentiated ductal carcinoma. "Single file" or targetoid patterns with extensive sclerosis mimicking invasive lobular carcinoma was common in lymphomas and was seen in one of the granulocytic sarcomas. In addition, two breast lymphomas, one of B-cell phenotype and the other of T-cell phenotype, showed frequent signet ring cells, which potentially could be confused with lobular carcinoma. Despite a number of recent articles on lymphomas of the breast, it appears that these tumors continue to be confused with carcinomas. Histologic features suggestive of lymphomatous involvement include absence of in situ carcinoma, frequent individual karyorrhectic cells, lymphoepithelial lesions, and cellular discohesiveness.
3,4-Epoxy-1-butene (EB) is the major mutagenic metabolite of butadiene (BD), an important industrial chemical classified as a probable human carcinogen. Although the mechanism of carcinogenicity of EB is not known, its reactions with nucleophilic sites of DNA giving pro-mutagenic lesions are likely to constitute the early crucial step in multistage carcinogenesis. This study was conducted to characterize the adducts formed from reactions of EB with the most nucleophilic DNA nucleobases, adenine (Ade) and guanine (Gua), as free nucleobases, 2'-deoxyribonucleosides and constituents of calf thymus DNA (CT DNA) in order to provide insight into the nature of DNA modification by EB. The adducts were isolated using HPLC separation coupled with diode array detection (DAD) and structurally characterized from their electronic, mass- and nuclear magnetic resonance spectra. Four EB-adenine products were identified as N-1-(2-hydroxy-3-buten-1-yl) adenine (EB-Ade I), N-1-(1-hydroxy-3-buten-2-yl) adenine (EB-Ade II), N-3-(2-hydroxy-3-buten-1-yl) adenine (EB-Ade III) and N-3-(1-hydroxy-3-buten-2-yl) adenine (EB-Ade IV). Two previously reported guanine adducts: N-7-(2-hydroxy-3-buten-1-yl) guanine (EB-Gua I) and N-7-(1-hydroxy-3-buten-2-yl) guanine (EB-Gua II) were also collected. The purified adducts were used as reference compounds to detect and quantitate the corresponding adduct species formed in calf thymus DNA incubated with EB. All six adducts were detected in treated DNA. The N-7 position of guanine was the most reactive in DNA followed by N-3 of adenine and N-1 of adenine. The formation of N-1 and N-3-adenine adducts (EB-Ade I, 1.2 +/- 0.36; EB-Ade II, 0.8 +/- 0.27; EB-Ade III, 2.7 +/- 0.38; EB-Ade IV, 5.9 +/- 0.68 nmol/micromol Ade) in CT DNA was approximately one-tenth that of EB-guanine adducts (50.7 +/- 2.37 and 47.9 +/- 3.6 nmol/micromol Gua, respectively). The N-1-EB-Ade adducts detected in this study are likely to be the precursors of previously reported N6-EB-adenine adducts (Koivisto et al., 1995) through Dimroth rearrangement. Since BD and EB induce significant numbers of point mutations at A:T base pairs, the EB-adenine adducts may represent important lesions involved in BD-induced mutagenesis and carcinogenesis.
Polycystic ovary syndrome (PCOS) is often associated with hyperinsulinaemia and peripheral insulin resistance. Whether the ovary is resistant to insulin is a matter of controversy. The aim was therefore to study the effect of insulin on lactate accumulation, an indicator of glucose metabolism, in granulosa-luteal cells from women with PCOS and from women with normal ovarian function. The cells were obtained from women undergoing clinical in-vitro fertilization-embryo transfer, either from patients with normal ovarian function and tubal or male infertility, or from women with PCOS, with or without tubal factor. The patients were down-regulated with buserelin and stimulated with urofollitrophin and human chorionic gonadotrophin (HCG). Follicle aspiration was performed under ultrasound guidance. Following oocyte recovery the granulosa-luteal cells were isolated, washed and cultured (2-3 x 10(4) viable cells/well) in serum-free Eagle's minimal essential medium for 48 h. After washing, the cells were then cultured in medium containing HCG (0.1-10 IU/ml) or insulin (0.05-0.5 microg/ml) for 24-48 h. Lactate accumulation in the media and cellular protein were analysed. Basal lactate accumulation did not differ in granulosa-luteal cells obtained from normal or PCOS ovaries, and averaged 46 and 49 nmol/g protein/24 h, respectively. A significant stimulation (40-60%) was obtained by HCG in both groups. Insulin caused a dose-dependent increase in lactate in granulosa-luteal cells obtained from normal ovaries (control: 45.5 +/- 6.3; insulin 0.5 microg/ml: 77 +/- 10 nmol/microg protein). Lactate accumulation in granulosa-luteal cells from PCOS ovaries was not altered in the presence of insulin. These results suggest that granulosa-luteal cell glucose metabolism is resistant to insulin in PCOS.
Nitrate increases the transcription of the two Arabidopsis thaliana nitrate reductase genes. We demonstrated previously that 238 and 330 bp of the 5' flanking regions, designated as NP1 and NP2, of the two nitrate reductase genes NR1 and NR2, respectively, are sufficient for nitrate-dependent transcription (Y. Lin, C.-F. Hwang, J.B. Brown, C.-L. Cheng [1994] Plant Physiol 106: 477-484). Here we identify the cis-acting elements of NP1 and NP2 that are necessary for nitrate-dependent transcription by linker-scanning (LS) analysis. In transgenic plants one LS mutant of NP1 and two LS mutants of NP2 exhibited significantly lower nitrate-induced reporter gene chloramphenicol acetyltransferase activity. To distinguish which of these three mutants lost nitrate inducibility, competitive reverse-transcriptase polymerase chain reaction was used to measure the chloramphenicol acetyltransferase mRNA levels before and after nitrate induction. The single LS mutant in NP1 lost its response to nitrate, whereas the two LS mutants in NP2 partially lost their response to nitrate. A 12-bp sequence is conserved between the NP1 site and the two NP2 sites. This sequence motif is also conserved in the 5' flanking regions of other nitrate-inducible plant genes. Gel mobility shift experiments indicate that these three regions bind to similar proteins. The binding is constitutive with respect to nitrate treatment and was observed in both nonphotosynthetic suspension cells and green leaves.
Light acts both directly as a signal and indirectly through photosynthesis to regulate the expression of genes encoding nitrate reductase (NR). Here, we report the isolation and characterization of a novel chlorate-resistant mutant that is defective in the regulation of NR gene expression. The response of NR2, but not NR1 or the gene encoding nitrate reductase (NiR), to light signals was impaired in this Arabidopsis mutant, designated cr88. In addition to NR2, the light regulation of the genes encoding the chlorophyll a/b binding protein (CAB) and the small subunit of ribulose bisphosphate carboxylase (RBCS) was also impaired in this mutant. These results suggest that the pathway through which light regulates the expression of NR2, CAB, and RBCS genes is different from those that regulate the expression of NR1 and NiR. An examination of the deetiolation process under different light spectrum showed that cr88 is defective in red light-mediated deetiolation. Complementation tests with various long hypocotyl (hy) mutants indicated that CR88 identifies a new HY locus. The possible functions of CR88 are discussed.
The goal of this study was to determine whether delivering transvenous defibrillation shocks, coordinated with the up/down-slope VF waveform patterns in the shocking lead, would improve the probability of successful defibrillation. Anesthetized swine (32-38 kg, n = 8) were implanted with an RV-->SVC + SQArray transvenous system to measure VF waveform patterns and to deliver shocks. The shocks were generated by a Cardiac Pacemakers Inc. biphasic waveform generator. Energy required for 50% success probability (E50) was determined using the multishock up-down protocol. VF was repeatedly induced and defibrillation shocks at E50 were given after 10 seconds. The defibrillation outcome, delivered energy (Ed), peak voltage (V), peak current (I), system impedance (Z) and VF waveform pattern at the time of shock were recorded and measured. Out of a total of 685 shocks, 324 (47%) succeeded and 361 (53%) failed. The Ed, V, I, and Z were similar for the two defibrillation outcome groups (success or failure). VF patterns were classified as high or low amplitude at the time of the shock based on the peak-to-peak amplitude of signals recorded between the shocking electrodes. Shocks that coincided with high amplitude VF patterns were further divided into shocks that occurred on the up-slope or on the down-slope. The probability of success when the E50 shocks were coincident with high or low amplitude fibrillation did not differ significantly (Student's t-test: 46% vs 48%. P = NS). However, during high amplitude fibrillation, shocks delivered on the up-slope were significantly more successful than those delivered on the down-slope (Chi-square: 67% vs 39%; P < 0.001). These results suggest that delivering defibrillation shocks during the up-slope of the high amplitude signal in the shocking lead may improve the probability of successful defibrillation of ICDs.
To investigate the hypothesis that Mycobacterium tuberculosis penetrates the alveolar epithelium by downregulating its barrier properties, we evaluated the interactions between M. tuberculosis and rat alveolar epithelial cell monolayers that are believed to share electrophysiologic properties of the human alveolar epithelium. Nonproteinaceous components of M. tuberculosis caused marked declines in electrical resistance and equivalent short-circuit current of the alveolar epithelial cell monolayers, indicating a reduction in the capacity to maintain tight intercellular junctions and to actively reabsorb sodium. M. tuberculosis elicited production of TNF-alpha mRNA and protein by alveolar epithelial cells, and the effects of recombinant TNF-alpha on the bioelectric properties of the alveolar epithelial paralleled those of M. tuberculosis. Furthermore, the effects of M. tuberculosis on alveolar epithelial resistance were abrogated by neutralizing anti-TNF-alpha antibodies. These results indicate that M. tuberculosis elicits production of TNF-alpha, which in turn reduces the bioelectric barrier properties of the alveolar epithelium. These findings provide insight into potential mechanisms by which M. tuberculosis establishes infection and disease in the lung.
A spatiotemporal ESR-CT study, rapid three dimensional ESR imaging by which distribution and metabolism of radicals in a small region in a living body can be followed, was carried out by intravenously administering spin-labeled polysaccharides to mice. The in vivo lifetime of spin-labeled hydroxyethylstarch (TEMPO-HES) was shorter than that of spin-labeled dextran (TEMPO-DX), suggesting that the clearance of their spins depended on the stability of the polymer chains to the hydrolysis. Spatiotemporal image at the kidney domain of a mouse showed that low molecular weight TEMPO-DX was excreted into the kidney and then was transferred to the bladder. Time dependence of the ESR intensity of TEMPO-HES at certain points in the liver domain had a maximum showing that TEMPO-HES was taken into the liver and then was decomposed there by the metabolism, but the rate of decrease in the intensity was different in each part in the liver because of the difference in metabolic function. The rate of decrease of TEMPO-DX in the blood was slowed by the prior administration of dextran, meaning that the intake of TEMPO-DX into the liver from the blood was suppressed by the dextran.
To estimate prevalence and incidence of adult Still's disease in Japan, and to describe the epidemiological features of the patients, a nationwide epidemiological survey was conducted in 1994. The study consisted of two questionnaires which were distributed to the heads of the relevant departments, randomly sampled, in hospitals throughout Japan. Following major epidemiological findings emerged from the study: (a) The total annual number of patients treated for adult Still's disease was estimated as 1,100 in 1993 in Japan. The estimated crude prevalence among those aged 16 years or older were calculated as 0.73 and 1.47 per 100,000 population for males and females, respectively, with the corresponding crude incidence rate of 0.22 and 0.34. (b) The sex ratio (female to male) of the reported patients was 2.1. This female predominance might be specific to Japan, though additional surveys in other countries will be warranted. The mean age of the patients was 38.1 years, and female patients tended to be older than male ones; 50% of the female patients aged 40 years or older, while so did only 28% of the male patients.
To disclose the association between smoking habits and lung cancer in Okinawa, Japan, we analyzed the data from a case-control study conducted from 1988 to 1991. The analysis, based on 333 cases and 666 age-, sex- and residence-matched population controls, provided the following major findings. (a) The odds ratios (ORs) for current smokers relative to nonsmokers were much greater for squamous cell carcinoma than for adenocarcinoma. The OR was 9.82 for squamous cell carcinoma and 2.18 for adenocarcinoma in males, 28.2 and 1.14, correspondingly, in females. (b) Males who quit smoking for 20 years or more demonstrated no elevated lung cancer risk. (c) Among male current smokers, the more the number of cigarettes smoked per day, the higher the lung cancer risk for both cell types, but particularly for squamous cell carcinoma. In contrast, deep smoke inhalation significantly increased the risk for adenocarcinoma in particular. (d) Okinawan brand cigarettes were more strongly associated with the risk, compared with other brand ones. This finding might partly explain the higher frequency of lung cancer in males with the relatively lower smoking rate in Okinawa.
OBJECTIVE: p16 and Rb protein were examined in primary astrocytomas in order to study the correlation between p16 and Rb proteins. METHODS: p16 and Rb pretoin were immunostained by SP immunohistochemical method in the sections of formalin fixed paraffin embedded tumor tissue from 102 patients with astrocytoma brain tumors. RESULTS: p16 and Rb protein were expressed in all low grade (WHO Grade I and II) astrocytomas but only in 48.1% and 57.4% astrocytomas of Grade III and Grade IV respectively, in which, 24/31 of Rb protein positive tumors showed null or low expression of p16 protein, while 19/23 of Rb negative tumors were associated with positive or high level expression of p16 protein. CONCLUSIONS: (a) p16 and Rb proteins were both involved in astrocytoma progression. (b) Negatively correlated pressionof p16 and Rb protein might be one of characteristics of malignant astrocytoma.
Fifty-two patients entered ICU after heart operation with cardiopulmonary bypass. These patients with ventilation atracurium (ATC) were given 3 micrograms-6 micrograms/kg/min for 5-48 hours. No complications happened. After ATC was used, the patients could open eyes, and cooperate with the doctor when they regain consciousness. The circulation index was normal. The limbs of these patients relaxed. In the 52 cases, 30 did't use sedatives, and 22 used senall dose of sedatives for sleep at night. We conclude that ATC does not require hepatic or renal function for termination of effect and action is terminated by the "Hoffman elimination" the ester hydrolysis. The time of effect is short and there is no cumulation effect. There is no vagal or ganglionic blocking activity, so ATC does not influence circulation and myocardial function. Maintenance relaxation keeps ventilation steady and oxygen supply normal.