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Biomedical subjects

Y F Li

Publications and source records attributed to Y F Li.

At least 19 recordsLinked to original sources

Expression and activity of matrix metalloproteinase 2 and 9 in human trophoblasts.

Indiscriminate invasion upon the endometrium by normal trophoblasts is strictly regulated unlike that by choriocarcinoma cells. In this study, we focused on the activity of matrix metalloproteinase (MMP)-2 and MMP-9 as parameters of invasion in normal human placenta. In situ hybridization (ISH), immunohistochemical staining (IH) and film in situ zymography (FIZ) were performed to identify cells having MMP-2 or MMP-9 expression and activity. Purified cytotrophoblasts (CTs) were used to examine the expression and activity of MMP-2 and MMP-9, and their invasive ability. In first trimester placental tissue, the MMP-2 expression was observed in extravillous trophoblasts (EVTs), and MMP-9 mainly in villous cytotrophoblasts (VCTs). FIZ revealed marked gelatinase activity in the EVTs which MMP-2 expression was observed in. In full-term placental tissue, the MMP-2 expressions was observed in the EVTs similar to that in first trimester, whereas the gelatinase activity in these cells was decreased or completely lost. Using purified CTs, the gelatinase activity was marked in early CTs, but not term CTs. Invasive ability of early CTs was inhibited by tissue inhibitor of metalloproteinase (TIMP)-2 and MMP-2 antibody in a dose dependent manner. These suggests that the invasive ability of trophoblasts may be regulated by the enzyme activity of gelatinases, especially MMP-2.

Adult↗

Identification of bulb from Fritillaria cirrhosa by PCR with specific primers.

Bulb of Fritillaria cirrhosa is an important traditional Chinese herbal medicine. According to the Chinese Pharmacopoeia (1995), it is commonly used as an antitussive and expectorant. Many young bulbs from species of Fritillaria are similar to those of F. cirrhosa, but they are different in price and quality. Therefore, there are many young bulbs from species of Fritillaria that could fake those of F. cirrhosa on the commercial market. The coding region of 5S-rRNA is highly conserved in higher eukaryotes. The 5S-rRNA spacer region sequences of F. thunbergii, F. pallidiflora, F. ussuriensi, F. delavayi, F.cirrhosa, F. anhuiensis, F. puqiensis were cloned by PCR with a pair of primers located within the conserved coding region. Based on sequences analyses of the 5S-rRNA spacer region from the 7 species, a specific sequence was found in F. cirrhosa. A pair of specific primers was designed for differentiating the bulbs of F. cirrhosa from each other by PCR. This result indicated that the method is rapid, more accurate and applicable in identification of the bulbs of F. cirrhosa at the DNA level.

Base Sequence↗

A pesticide runoff model for simulating runoff losses of pesticides from agricultural lands.

An integrated modeling system was developed to predict runoff losses of pesticides from agricultural lands. The system is an integration of a mathematical model, a database system, and a geographic information system. Information on soil type, land use, land slope, watershed boundaries, precipitation, pesticide usage, as well as physical and chemical properties of pesticides have been input to a GIS, managed through a database, and used for further modeling studies. The modeling outputs were in turn put into the database, such that runoff patterns along with pesticides losses could be further simulated by using a database management system. The final results could then be visualized through GIS. The developed modeling system was applied to the Kintore Creek Watershed, Ontario, Canada, for simulating losses of atrazine from agricultural lands. A water quality monitoring project was carried out from 1988 to 1992 in the watershed to detect conditions of surface water pollution due to the use of pesticides. The modeling outputs were verified through the monitoring data, demonstrating reasonable prediction accuracy. The result indicated that the model provides an effective means for forecasting pesticide runoff from agriculture lands.

Agriculture↗

The transport of beta-hexachlorocyclohexane to the western Arctic Ocean: a contrast to alpha-HCH.

A large database for alpha-hexachlorocyclohexane (alpha-HCH), together with multimedia models, shows this chemical to have exhibited classical 'cold condensation' behavior. The surface water of the Arctic Ocean became loaded between 1950 and 1990 because atmospheric transport of alpha-HCH from source regions to the Arctic was rapid and because alpha-HCH partitioned strongly into cold water there. Following emission reductions during the 1980s, alpha-HCH remained trapped under the permanent ice pack, with the result that the highest oceanic concentrations in the early 1990s were to be found in surface waters of the Canada Basin. Despite a much stronger partitioning into water than for alpha-HCH, beta-HCH did not accumulate under the pack ice of the Arctic Ocean, as might be expected from the similar emission histories for the two chemicals. Beta-HCH appears to have loaded only weakly into the high Arctic through the atmosphere because it was rained out or partitioned into North Pacific surface water. However, beta-HCH has subsequently entered the western Arctic in ocean currents passing through Bering Strait. Beta-HCH provides an important lesson that environmental pathways must be comprehensively understood before attempting to predict the behavior of one chemical by extrapolation from a seemingly similar chemical.

Arctic Regions↗

Crystallization and preliminary X-ray analysis of Citrobacter amalonaticus methylaspartate ammonia lyase.

Methylaspartate ammonia lyase (MAL) catalyses the reversible alpha,beta-elimination of ammonia from L-threo-(2S,3S)-3-methylaspartic acid to give mesaconic acid. Crystals of Citrobacter amalonaticus MAL have been obtained by the hanging-drop method of vapour diffusion using ammonium sulfate as the precipitant. Three crystal forms were obtained from identical crystallization conditions, two of which (forms A and B) diffract to high resolution, whilst the third form diffracted poorly. Crystals of form A diffract to beyond 2.1 A and have been characterized as belonging to one of the enantiomorphic space groups P4(1)22 or P4(3)22, with unit-cell parameters a = b = 66.0, c = 233.1 A, alpha = beta = gamma = 90 degrees and a monomer in the asymmetric unit. Crystals of form B diffract to beyond 1.5 A and belong to space group C222, with unit-cell parameters a = 128.3, b = 237.4, c = 65.8 A, alpha = beta = gamma = 90 degrees and a dimer in the asymmetric unit. Determination of the structure of MAL will be an important step in resolving current conflicts concerning the enzyme mechanism which differ between one which places MAL as a member of the superfamily of ammonia lyases whose catalytic activity requires a cofactor formed by post-translational modification of the enzyme and another which links MAL to the enolase superfamily.

Ammonia-Lyases↗

Total internal reflected resonance light scattering determination of chlortetracycline in body fluid with the complex cation of chlortetracycline-europium-trioctyl phosphine oxide at the water/tetrachloromethane interface.

A highly selective method of chlortetracycline (CTC) is proposed on the basis of the measurements of total internal reflected resonance light scattering (TMR-RLS) at water/tetrachloromethane (H20/CCl4) interfaces. In the pH range of 7.54-8.14, the interaction of the binary complex of Eu(III)/CTC in the presence of trioctyl phosphine oxide (TOPO) occurs at the H20/CCl4 interface, resulting in greatly enhanced TIR-RLS signals with the maximum peak located at 340 nm. The enhanced TIR-RLS intensity is in proportion to the CTC concentration in the range 0.98 to approximately 20.0 x 10(-7) mol/L. The limit of detection is 9.8 x 10(-9) mol/L. Synthetic samples and body fluid samples including human urine, human serum, and fresh milk were determined with the recovery of 95.4-106.4% and RSD of 2.9-3.9%.

Animals↗

Physical activity and breast cancer risk in hispanic and non-hispanic white women.

To investigate breast cancer risk in Hispanic and non-Hispanic White women, the authors conducted a population-based case-control study of New Mexican women during 1992-1994 using incident breast cancer cases aged 35-74 years and frequency-matched controls selected using random digit dialing. Activity type and weekly duration of usual nonoccupational physical activity were used to calculate weekly metabolic equivalent (MET)-hours of total and vigorous physical activity (> or =5 METs). Conditional logistic regression models were fitted to estimate the relative risk of breast cancer for levels of physical activity and to assess the difference in effects by ethnicity, body mass index, energy intake, and menopausal status. Vigorous physical activity was associated with reduced breast cancer risk in both Hispanic and non-Hispanic White women. Women in the highest category of vigorous activity had lower risk of breast cancer (adjusted odds ratio = 0.34, 95% confidence interval: 0.22, 0.51 for Hispanic; adjusted odds ratio = 0.60, 95% confidence interval: 0.41, 0.89 for non-Hispanic White women) compared with women reporting no vigorous physical activity. Both pre- and postmenopausal Hispanic women showed decreasing risk with increasing level of activity. Physical activity was protective only among postmenopausal non-Hispanic White women. The effects of physical activity were independent from reproductive factors, usual body mass index, body mass index at age 18, adult weight gain, and total energy intake.

Adult↗

In vivo mechanism-based inactivation of S-adenosylmethionine decarboxylases from Escherichia coli, Salmonella typhimurium, and Saccharomyces cerevisiae.

S-adenosylmethionine decarboxylase (AdoMetDC), a key enzyme in the biosynthesis of spermidine and spermine, is first synthesized as a proenzyme, which is cleaved posttranslationally to form alpha and beta subunits. The alpha subunit contains a covalently bound pyruvoyl group derived from serine that is essential for activity. With the use of an Escherichia coli overexpression system, we have purified AdoMetDCs encoded by the E. coli, Saccharomyces cerevisiae, and Salmonella typhimurium genes. Unexpectedly we found by mass spectrometry that these enzymes had been modified posttranslationally in vivo by a mechanism-based "suicide" inactivation. A large percentage of the alpha subunit of each enzyme had been modified in vivo to give peaks with masses m/z = 57 +/- 1 and m/z = 75 +/- 1 daltons higher than the parent peak. AdoMetDC activity decreased markedly during overexpression concurrently with the increase of the additional peaks for the alpha subunit. Sequencing of a tryptic fragment by tandem mass spectrometry showed that Cys-140 was modified with a +75 +/- 1 adduct, which is probably derived from the reaction product. Comparable modification of the alpha subunit was also observed in in vitro experiments after incubation with the substrate or with the reaction product, which is consistent with the in vitro alkylation of E. coli AdoMetDC reported by Diaz and Anton [Diaz, E. & Anton, D. L. (1991) Biochemistry 30, 4078-4081].

Adenosylmethionine Decarboxylase↗

The distribution patterns of trace elements in the brain and erythrocytes in a rat experimental model of iodine deficiency.

Cretinism is a disease characterized by neurological defects associated with severe iodine deficiency. In a rat model of severe iodine deficiency, we investigated the distribution pattern of trace elements (iodine [I], selenium [Se], and bromine [Br] in brain tissue samples; potassium [K], calcium [Ca], manganese [Mn], iron [Fe], copper [Cu], zinc [Zn], rubidium [Rb], and lead [Pb] in erythrocytes) after supplementing the rats with I and/or Se. Neutron activation analysis, proton induced x-ray emission and x-ray fluorescence were used. The serum levels of total and free thyroxine (T4, FT4), and of total, free, and reverse triiodothyronine (T3, FT3, rT3, respectively) were assessed by radioimmunoassay. The results were statistically evaluated by one-way analysis of variance and bivariate correlation. The study indicated that the levels of T4, FT4, and rT3 increased in the serum of iodine-deficient rats supplemented with I or I + Se. In the same animals, we documented alterations of the content of Br in the brain, and of Zn, Mn, Cu, and Rb in erythrocytes, whereas the brain content of I and Se was unchanged. Thus, I and I + Se supplementation improves thyroid hormone metabolism but affects the content of selected trace elements in erythrocytes and of Br in the brain. The data stimulate further clarification of the role of trace elements in the central nervous system.

Animals↗

Ectopic expression of TATA box-binding protein induces shoot proliferation in Arabidopsis.

The TATA box-binding protein (TBP) is an essential component of transcription initiation complexes of all three eukaryotic RNA polymerases. Increasing evidence suggests that the TBP activity and availability may be regulated to precisely control gene transcription and play an important role in cell type-specific regulation. Arabidopsis TBP-2 is up-regulated in apical shoot tissues. Overexpression of TBP-2 in transgenic Arabidopsis induces apical shoot proliferation. The expression of some shoot meristem regulatory genes is altered. These data suggest that the TBP gene dosage and/or expression level may play an important role in controlling shoot production and plant morphology.

Arabidopsis↗

Use of an in vitro immunoselected tumor line to identify shared melanoma antigens recognized by HLA-A*0201-restricted T cells.

An immunoselected melanoma cell line that had lost expression of the dominant melanoma antigens MART-1 and gp100 was generated in an attempt to identify previously unknown tumor antigens. After repeated stimulation with the autologous immunoselected tumor line, a number of HLA-A*0201-restricted T-cell clones were established from the peripheral blood of a single melanoma patient. One T-cell clone (C-22) recognized 14 of 16 HLA-A2+ melanoma cell lines, as well as HLA-A2+ melanocytes but recognized neither HLA-A2+ fibroblasts nor autologous B cells. Screening of an autologous cDNA library resulted in the isolation of a transcript identical to an entry in the expressed sequence tag database. Northern blot analysis revealed that this gene was expressed in most melanoma cell lines and melanocytes but not in normal tissues. The peptide epitope (AMF-GREFCYA) recognized by clone C-22 was identified based on studies of the recognition of truncated cDNAs and the use of the consensus HLAA*0201 binding motif. A second T-cell clone (C-29) was found to recognize a new tyrosinase-related protein 2 epitope (455-463; YAIDLPVSV) in an HLA-A*0201-restricted manner. Together, these results provide additional targets that can be used for the development of immunotherapeutic protocols in HLA-A2+ melanoma patients and demonstrate the utility of immunoselected tumor lines for the identification of new melanoma antigens.

Amino Acid Sequence↗

Altered mRNA expression of sialyltransferase in squamous cell carcinomas of the cervix.

OBJECTIVE: Increased sialylation has been reported in various kinds of cancers, but to date, sialylation of cervical carcinoma has never been evaluated. This study of the changes in messenger ribonucleic acid (mRNA) expression of the four sialyltransferases (ST3Gal I, ST3Gal III, ST3Gal IV, and ST6Gal I) in a normal cervix and that with FIGO stage IB1 squamous cell carcinoma was undertaken to assess the extent of sialylation associated with establishment of the carcinoma. METHODS: Alterations in ST mRNA expression in FIGO IB1 cervical cancer (n = 30) and normal cervixes (n = 30) were examined by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: ST6Gal I expression was enhanced in squamous cell carcinoma of the cervix (P = 0.026, Mann-Whitney U test), but mRNA expression from the other three STs (ST3Gal I, ST3Gal III, and ST3Gal IV) was significantly down-expressed in squamous cell carcinoma of the cervix compared to the normal cervix (P = 0.003, P < 0.001, and P = 0.001, respectively). High ST6Gal I expression was associated with more invasive properties of cervical cancer, such as deep stromal invasion, lymph or vascular space involvement, and poor differentiation (P = 0.010, P < 0.001, P < 0.001, respectively). CONCLUSIONS: A combination of enhanced ST6Gal I mRNA expression and decreased mRNA expression from ST3Gal I, ST3Gal III, and ST3Gal IV might be important in cervical cancer. Future studies will investigate whether RT-PCR detection of the expression of these enzymes can be helpful for prognostic purposes.

Carcinoma, Squamous Cell↗

Determination of proteins by their enhancement of resonance light scattering by fuchsine acid.

Based on the measurement of the enhancement of resonance light scattering (RLS) of fuchsine acid (FSA) by proteins, a novel sensitive assay of proteins in body fluid samples has been developed. Proteins, including bovine serum albumin (BSA), human serum albumin (HSA), pepsin (Pep), alpha-chymotrypsin (Chy), lysozyme (Lys), and cellulase (Cel), can bind to fuchsine acid (FSA), resulting in enhanced RLS signals at 277.0 nm. Linear relationships between the enhanced RLS intensity and the protein concentration were measured at different concentration of FSA, and the limits of detection for BSA, HSA, and Lys were found to lie in the nanogram range.

Animals↗

Gridded usage inventories of technical hexachlorocyclohexane and lindane for China with 1/6 degrees latitude by 1/4 degrees longitude resolution.

China banned the use of technical HCH (BHC: 1,2,3,4,5,6-hexachlorocyclohexane) in 1983; lindane has been used in this country since 1991. The total production was around 4 million t for technical HCH from 1952 to 1984, and 11,400 t for lindane between 1991 and 2000. Though the total produced technical HCH was considered to be used in China, only 3,200 t of lindane were used between 1991 and 2000 with the rest for export or on unused stack. Annual usage of these two compounds was located into each province first and then broken for different crops. Inventories of gridded usage of both technical HCH and lindane with 1/6 degrees by 1/4 degrees latitude/longitude resolution have been created by different gridded cropland data sets as surrogate data. The intensive use of technical HCH on croplands was concentrated in the southeastern part of China, but the use of lindane was concentrated in the northern part of China.

Agriculture↗

Papain immobilization on a nitrilon fibre carrier containing primary amine groups.

A carrier containing primary amine groups was synthesized by the reaction between nitrilon fibre and diethylenetriamine, and papain was immobilized on this kind of carrier by using glutaraldehyde as a coupling agent. The factors involved with the activity recovery of the immobilized papain and the enzymic properties of the resulting immobilized papain were studied in comparison with free papain, for which casein was chosen as a substrate. The results show that the activity recovery of immobilized papain can reach 47.3%, and that the immobilized papain exhibits better environmental adaptability and reusability than free papain.

Amines↗

Comparative efficacy of fibrinogen and platelet supplementation on the in vitro reversibility of competitive glycoprotein IIb/IIIa (alphaIIb/beta3) receptor-directed platelet inhibition.

BACKGROUND: Platelet surface glycoprotein (GP) IIb/IIIa (alphaIIb/beta(3)) receptor inhibition, by preventing fibrinogen binding and platelet aggregation, concomitantly attenuates arterial thrombotic capacity and impairs protective hemostasis, 2 divergent platelet-dependent processes. Because the currently available Food and Drug Administration-approved small molecule GP IIb/IIIa receptor antagonists are considered "competitive" inhibitors and because there is limited information on the reversibility of platelet inhibition by fibrinogen or platelet supplementation, the following series of in vitro experiments were performed. METHODS AND RESULTS: Washed platelets from 24 healthy volunteers were suspended in Tyrodes buffer and incubated with achievable (in vivo) steady-state concentrations of either tirofiban or eptifibatide before activation with TRAP (thrombin receptor agonist peptide) (15 micromol/L). Platelet aggregation was inhibited by 40% to 50%, but reversal was achieved by fibrinogen supplementation in a concentration-dependent manner. In a separate series of in vitro experiments, platelet inhibition exceeding 90% was established with tirofiban (average concentration 9.28 microg/L) and eptifibatide (average concentration 95.4 microg/L). Recovery of platelet aggregation to at least 50% was achieved after the addition of fibrinogen (0.76-0.80 g/L), platelets (2.4 x 10(11)/L), or their combination. There was an inverse relationship between plasma baseline fibrinogen and the amount of supplemental fibrinogen required to restore platelet aggregability (r = -0.60, P <.01). CONCLUSIONS: The reversibility of GP IIb/IIIa-directed platelet inhibition is influenced by cell surface receptor availability and the intrinsic pharmacodynamic mechanism of action. Fibrinogen supplementation with fresh frozen plasma or cryoprecipitate either alone or in combination with platelet transfusion, represents an important and readily available treatment consideration for restoring hemostatic potential and managing major hemorrhagic complications associated with the administration of small molecule competitive GP IIb/IIIa receptor antagonists.

Adult↗

Blunted nitric oxide-mediated inhibition of renal nerve discharge within PVN of rats with heart failure.

We have demonstrated a decreased neuronal nitric oxide (NO) synthase (nNOS) message in the hypothalamus of rats with heart failure (HF). Subsequently, we have demonstrated that NADPH diaphorase (a commonly used marker for nNOS activity) positive neurons are decreased in paraventricular nucleus (PVN) of rats with coronary artery ligation model of HF. The goal of the present study was to examine the influence of endogenous NO within the PVN on renal sympathetic nerve discharge (RSND) during HF. In alpha-chloralose- and urethane-anesthetized rats, an inhibitor of NO synthase, N(G)-monomethyl-L-arginine (L-NMMA) microinjected into the PVN (50, 100, and 200 pmol in 50-200 nl) produced a dose-dependent increase in RSND, blood pressure, and heart rate in control and HF rats. These responses were attenuated in rats with HF compared with control rats. On the other hand, the NO agonist, sodium nitroprusside, microinjected in PVN produced a dose-dependent decrease in RSND and blood pressure in control and HF rats. These responses were less in rats with HF compared with control rats. These data suggest that the endogenous NO-mediated effect within the PVN of HF rats is less potent in suppressing RSND compared with control rats. These data support the conclusion that the NO system within the PVN involved in controlling autonomic outflow is altered during HF and may contribute to the elevated levels of renal sympathoexcitation commonly observed in HF.

Animals↗