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Biomedical subjects

L Xiao

Publications and source records attributed to L Xiao.

At least 19 recordsLinked to original sources

Cyclic guanosine 3',5'-monophosphate mediates 3-morpholinosydnonimine-induced inhibition of human natural killer cells.

Nitric oxide (NO) donors were used to investigate the effect of NO on and the role of cyclic GMP in the regulation of human natural killer (NK) cell function. NO-producing drugs, molsidomine and its metabolite 3-morpholinesydnonimine (SIN-1), inhibited NK cell-mediated cytotoxicity significantly at 0.04-5 mM. At 1 mM, SIN-1 completely inhibited NK cell activity while molsidomine decreased NK cell-mediated cytolysis by 35% of the control value. These data suggest that NO from exogenous NO-donors may down-regulate NK cell cytotoxic function. The stimulatory effect of interferon-gamma (IFN-gamma) on human NK cell-mediated killing could not overtake the NK cell inhibition induced by the NO releasing drugs, indicating different modes of action for IFN-gamma and SIN-1. The results in the present study also showed that SIN-1 (1 mM) stimulated cyclic GMP production 37-fold in NK cells. In the presence of 0.5 mM IBMX, a phosphodiesterase inhibitor, the increase in cyclic GMP was even more pronounced, demonstrating a relation between cyclic GMP stimulation and NK cell inhibition by SIN-1. Further evidence for mediation via cyclic GMP was provided by the finding that methylene blue (20 microM), an inhibitor of soluble guanylate cyclase, decreased both the inhibition of SIN-1-induced NK cell cytotoxicity as well as cyclic GMP formation. Moreover, membrane-penetrating cyclic GMP and its analogues inhibited NK cell-mediated cytolysis significantly. Molsidomine was without effect on cyclic GMP levels. Our data indicate that cyclic GMP may play a role in human NK cell regulation and suggest that the inhibitory effect of cGMP may be elicited by NO.

1-Methyl-3-isobutylxanthine

Significant induction of spermidine/spermine N1-acetyltransferase without cytotoxicity by the growth-supporting polyamine analogue 1,12-dimethylspermine.

The superinduction of the polyamine catabolic enzyme spermidine/spermine N1-acetyltransferase (SSAT) has been implicated in the cell type-specific cytotoxic activity of some polyamine analogues. We now report that one polyamine analogue, 1,12-dimethylspermine (DMSpm), produces a large induction of SSAT with no significant effects on growth in the human large cell lung carcinoma line, NCl H157. This cell line has been demonstrated to respond to other analogues with SSAT superinduction and cell death. Treatment of the lung cancer cell line with DMSpm produces a rapid increase in SSAT activity and a near complete depletion of the natural polyamines. Additionally, DMSpm supports cell growth in cells which have been depleted of their natural polyamines by the ornithine decarboxylase inhibitor, 2-difluoromethylornithine. The current results suggest that significant induction of SSAT can occur in the absence of cytotoxicity when the inducing polyamine analogue can support growth and that increased SSAT activity alone is not sufficient for cytotoxicity to occur.

Acetyltransferases

Nitric oxide synthase pathway may mediate human natural killer cell cytotoxicity.

The present study provides evidence that the human natural killer (NK) cell effector mechanism causing target cytolysis has a requirement for L-arginine. In a deficient medium (DM) containing only salts, buffer system and glucose, NK cell-mediated cytotoxicity was found to decrease by 70% as compared to that obtained in a complete medium (CM). However, adding L-arginine to such DM could restore the activity of NK cells to the normal level. Many other components of CM, such as serum, glutamine and vitamins did not improve NK cell-mediated killing in DM. When all amino acids except L-arginine were added to DM only a partial recovery of NK cell functional cytolysis was seen. L-arginine enhanced the NK cell activity in a dose-dependent manner. Additionally, the inhibitor of both inducible and constitutive nitric oxide synthase, N-monomethyl-L-arginine (L-NMMA) inhibited NK cytolytic activity in DM supplemented with L-arginine indicating participation of nitric oxide (NO). The results also show that the stimulatory effect of L-arginine on human NK cell-mediated cytotoxicity was accompanied by an increase in NO formation as determined by accumulation of nitrite and citrulline. L-NMMA gave a dose-dependent reduction in NO generation as well. The nitrite and citrulline production dose-dependently correlated with not only the concentration of L-arginine in the cultivation medium, but also the enhanced NK cell-mediated cytolysis. Taken together, these findings could define a L-arginine/NO-linked effector mechanism in human NK cells. Nitrite and citrulline were not formed when NK cell-mediated target cell killing took place in a L-arginine-free DM supplemented with additives. Thus, it appears as if human NK cells may cause target cell killing via both NO-dependent and -independent processes.

Arginine

[Changes of HDL-receptor of hepatic cell membrane in rabbit model of calcium bilirubinate cholangiolithiasis].

HDL-receptor of the hepatic cell membrane and Fch level of the hepatic tissue in the rabbit model of calcium bilirubinate cholangiolithiasis were examined. The results showed: (1)the HDL-receptors' Bmax of the biliary obstruction and infection (BOI) and biliary obstruction (BO) groups decreased during the 1st week after operation (P < 0.01); they were only 69.09% and 49.09% of that in the control group respectively, and the lowered level continued until the 6th week; the Kd values of BOI and BO groups were not different from that of the control group during the 1st week, but they were significantly higher in the 6th week after operation (P < 0.01); (2)the Fch levels of the hepatic tissue were slightly decreased in all experimental groups (P < 0.05). We concluded that under the conditions of biliary obstruction and infection, there was a disorder of the HDL receptors of hepatic cell membrane. At the early stage of biliary obstruction and infection, the receptors decreased in number, and at the later stage, the affinity changed and less HDL entered into the hepatic cell. Then the precurser cholesterol pool for BA biosynthesis narrowed. Thus, BA secreted from the liver cells into genetic elements possibly promote the formation of the calcium bilirubinate gallstones.

Animals

Cytoplasmic Ca2+ inhibits the glucose transporter of human erythrocytes.

The effect of Ca2+ on the glucose transporter of human erythrocytes was investigated. The results showed that extracellular Ca2+ had no effect. But, the glucose transport of erythrocytes was markedly inhibited due to the increase in cytoplasmic Ca2+ concentration by addition of ionophore A23187. The Ca2+ inhibition exhibited a dose-dependent manner with an apparent half maximal concentration of 250 microM and could not be recovered by 10 mM EGTA. Unlike Ca2+, Mg2+ did not affect the glucose transporter.

Calcimycin

Quantitative microdialysis determination of extracellular striatal dopamine concentration in male and female rats: effects of estrous cycle and gonadectomy.

Sex differences in basal extracellular striatal dopamine concentrations in gonadectomized male and female rats have been reported previously. In the current experiment, estrous cycle-dependent variation, sex differences and the effect of gonadectomy on extracellular striatal dopamine concentrations were determined using quantitative microdialysis. Female rats were found to have significantly higher extracellular striatal dopamine concentrations in proestrus and estrus than in diestrus or after ovariectomy. In contrast, castration of male rats had no effect on extracellular striatal dopamine concentrations. Thus, endogenous ovarian hormones, but not testicular hormones, modulate extracellular striatal dopamine concentrations in rats.

Animals

[Alterations of oxygen free radicals in rabbit models and its relation to formation of pigment gallstones].

The experimental rabbits were divided into three groups. In the first group the common bile ducts were loosely ligated and contaminated by E. coli (BOI); in the second group the common bile ducts were only loosely ligated (BO); and the third group was the control group (Con). Alterations of oxygen free radicals were studied in all these models. The result showed that alterations of the partial hepatic oxygen free radicals (OFR), the hepatic prostaglandin E2 (PGE2) and the pigment gallstones were similar. There was a positive correlation among them (P < 0.05). In group BOI, these indexes were higher than in group BO. It is suggested that oxygen free radicals are possibly responsible for the formation of pigment gallstones through the pathway from oxygen free radicals to promote the production of prostaglandin E2, glycoprotein and pigment gallstones at last E. coli may aggravate the formation of pigment gallstones by increasing hepatic oxygen free radicals.

Animals

Steady-state messenger RNA and activity correlates with sensitivity to N1,N12-bis(ethyl)spermine in human cell lines representing the major forms of lung cancer.

Our previous results from a limited number of cell lines have suggested that the bis(ethyl)polyamine analogues exert a phenotype-specific response in human lung cancer cells. In the present study, we have extended this work to analyze the response of the 4 major forms of human lung cancer to the polyamine analogue N1,N12-bis(ethyl)spermine (BESpm). The results suggest that non-small cell phenotypes are much more sensitive to the cytotoxic effects of BESpm than the small cell lung carcinoma phenotype. Further, there appears to be a positive association between the level of induction of the polyamine catabolic enzyme spermidine/spermine N1-acetyltransferase (SSAT) in response to the analogue and the kinetic response of cells. Specifically, cells in which SSAT activity is highly induced by BESpm are killed by the compound. Although induction of SSAT appears to occur at both the level of increased steady-state mRNA and enzyme activity, SSAT activity appears to be a better indicator of cell sensitivity to BESpm than SSAT mRNA levels. These results have significance both for the potential use of polyamine analogues in treating specific forms of human lung cancer and for understanding the regulation of SSAT at the molecular level.

Acetyltransferases

Structure of the human spermidine/spermine N1-acetyltransferase gene (exon/intron gene organization and localization to Xp22.1).

The super induction of spermidine/spermine N1-acetyltransferase (SSAT), has been implicated in the cytotoxic response of human solid tumors to the bis(ethyl)polyamines. The SSAT response is a phenotype specific response and is modulated at the level of increased steady-state mRNA levels and enzyme protein. The human genomic region (4,095 bases) containing the coding sequence of SSAT has been cloned and localized to the Xp22.1 region. Primer extension analysis indicates the transcription of SSAT starts 179 bases upstream from the translational start site and appears to be under the control of a "TATA-less" promoter. The availability of this human clone will facilitate the direct functional examination of the SSAT gene.

Acetyltransferases

Molecular dynamics simulation of bovine prothrombin fragment 1 in the presence of calcium ions.

Early solvation-induced structural reorganization of calcium prothrombin fragment 1 is simulated with molecular dynamics. Initial coordinates are those of the 2.2-A resolution crystal structure [Soriano-Garcia, M., Padmanabhan, K., de Vos, A. M., & Tulinsky, A. (1992) Biochemistry 31, 2554-2556]. The molecular dynamics code AMBER, appropriately modified to include long-range (less than or equal to 22.0 A) ionic forces, was employed. The solution structure appears to equilibrate within 100 ps. Although minor changes are seen in various structural domains, the early solution structure basically maintains an intricate network of nine gamma-carboxyglutamic acid (Gla) residues encapsulating seven calcium ions. However, the Gla domain moves with respect to the kringle domain. This motion is mainly due to the movement of Ser34-Leu35 that appears to be a flexible hinge between the domains. The N-terminus of Ala 1 is in a tightly bound complex with three Gla residues that remains stable in the solution structure when the long-range electrostatic cutoff is employed and the near planar alignment of the seven calcium ions is only slightly distorted. The simulation structure is discussed in terms of experiments that studied calcium ion-induced quenching of the intrinsic fluorescence, protection of the N-terminal amino group from acetylation by calcium ions, chemical modification of the N-terminus to a trinitrophenyl derivative, and the possibility of a calcium-binding site(s) in the kringle domain.

Amino Acid Sequence

Infectivity of Moniezia benedeni and Moniezia expansa to oribatid mites from Ohio and Georgia.

Six species of oribatid mites belonging to four families and five genera were successfully infected by M. benedeni. They were Scheloribates laevigatus (Scheloribatidae), Exoribatula sp. cf. biundatus (Scheloribatidae), Xylobates capucinus (Haplozetidae) and Zygoribatula undulata (Oribatulidae) from Ohio, and Galumna ithacensis (Galumnidae) and Scheloribates lanceoliger (Scheloribatidae) from Georgia. Cysticercoids were also recovered from S. laevigatus infected with M. expansa. Among the six species of mites, S. laevigatus is the major intermediate host of M. expansa and M. benedeni worldwide, while the other five species have not been previously reported as transmitters of Moniezia spp. This is also the first report of any species from the genera Exoribatula and Xylobates as intermediate hosts of tapeworms. Different species of oribatid mites varied in their efficiency as intermediate hosts of M. benedeni. Moniezia benedeni had higher infection rate and infection intensity in S. laevigatus than in other species. Development in S. laevigatus was also more rapid than in the other species. Mite size was only one of the determinants of intermediate host efficiency. Storage at 4 degrees C for 1 to 5 weeks reduced the viability of M. benedeni eggs. Stored eggs had lower infection rates and infection intensity and slower development in S. laevigatus. They also had lower infection rates in X. capucinus and Exoribatula sp. cf. biundatus.

Animals

Effects of clinically apparent and subclinical Ostertagia ostertagi infections on nitrogen and water metabolism in calves.

Four balance trials were conducted in 3 groups of 5 calves each at 0, 4, 8, and 14 weeks after initial inoculation with Ostertagia ostertagi. Group-1 calves were inoculated with 100,000 third-stage larvae (L3) of O ostertagi/wk for 14 weeks. Group-2 calves were inoculated with 10,000 L3/wk for 14 weeks, and group-3 calves were not inoculated. Effects of infection on apparent digestibilities of dry matter and nitrogen, and balances of nitrogen and water were evaluated. Neither clinically apparent (group 1) nor subclinical (group 2) O ostertagi infections had observable effects on the apparent digestibility of dry matter. Subclinical infection also had no significant effects on nitrogen digestibility or nitrogen and water balance. Clinically apparent infection, however, decreased the apparent digestibilities of nitrogen significantly (P < 0.05) at 4, 8, and 14 weeks after inoculation, and decreased nitrogen balance at 4 and 8 weeks after inoculation. Group-1 calves also had lower water intake, fecal-water excretion, and apparent water balance, but higher urinary water output at 4, 8, and 14 weeks after inoculation.

Animals

Nutritional and pathophysiologic effects of clinically apparent and subclinical infections of Ostertagia ostertagi in calves.

Nutritional and physiologic effects of clinically apparent and subclinical Ostertagia ostertagi infections were studied in 3 groups of 5 calves each. Group-1 calves were inoculated with 100,000 Ostertagia ostertagi third-stage larvae (L3)/calf/wk for 14 weeks. Group-2 calves were inoculated with 10,000 L3/calf/wk for 14 weeks, and group-3 calves were no inoculated. Calves in group 1 had decreased dry matter intake and feed utilization from 4 weeks after initial inoculation. Group-2 calves had no changes in dry matter intake, but had decreased feed utilization at 12 and 14 weeks. Calves with clinically apparent infections (group 1) lost a mean weight of 11.8 kg, whereas calves with suclinical infections (group 2) lost a mean of 46.6 kg, and control calves lost a mean of 60.7 kg. Calves with O ostertagi infections (group 1 and 2) also had decreased carcass quality at slaughtering, which was reflected in decreased dressing weights and increased water-holding capacity of the rib-eye muscle. Calves in groups 1 and 2 also had lower carcass yield and rib-eye muscle weight, and group-1 calves had decreased protein content. Results of hematologic, pathologic, parasitologic, and clinical examinations mirrored nutritional changes.

Animal Nutritional Physiological Phenomena

Effects of Ostertagia ostertagi infection on secretion of metabolic hormones in calves.

Effects of Ostertagia ostertagi infection on secretion of insulin, pancreatic glucagon, cortisol, gastrin, and pepsinogen were studied in calves inoculated with 100,000 (group 1) or 10,000 (group 2) O ostertagi infective larvae weekly for 14 weeks. Plasma insulin concentrations in both inoculated groups were lower than those in a non-infected (group 3) control group. The differences between group 1 and group 3 were significant (P < 0.05) at 2 and 12 weeks after initial inoculation. Plasma pancreatic glucagon and cortisol concentrations of groups 1 and 2 did not differ significantly from those of the control group, although plasma pancreatic glucagon concentration was consistently lower in group-1 calves from 4 weeks to end of the study. Plasma pepsinogen and serum gastrin concentrations also increased significantly (P < 0.05) in both groups that received inoculations. We concluded that decreased plasma insulin concentrations are contributory to changes in postabsorptive protein metabolism, and that serum gastrin concentrations are more representative of the pathologic changes in the abomasum than are plasma pepsinogen concentrations.

Animals

Characterization of a full-length cDNA which codes for the human spermidine/spermine N1-acetyltransferase.

Spermidine/spermine N1-acetyltransferase is the rate-limiting enzyme in the catabolism of cellular polyamines. Using a combination of cDNA library screening and anchored PCR methodologies, a full length cDNA designated AP3/F7 corresponding to the human SSAT was cloned using RNA from the human large cell undifferentiated lung carcinoma line NCI H157. The resulting cDNA clone is 1,060 base pairs with a 513 base open reading frame coding for a 171 amino acid protein, with a predicted subunit molecular weight of 20,023. The 5' non-coding region of AP3/F7 is 165 bases and the 3' untranslated region is 382 bases with a polyadenylation site 20 bases 5' to the poly(A) tail. This full length cDNA should be an aid in the study of the regulation of spermidine/spermine N1-acetyltransferase expression and the significance of the acetyltransferase in polyamine metabolism.

Acetyltransferases

Photoaffinity polyamines: sequence-specific interactions with DNA.

ANB-spermine is a photoaffinity analog of the naturally-occurring polyamine, acetylspermine. ANB-spermine was used to determine its binding sites on naked double stranded DNA, at the nucleotide level, using a modification of the primer extension technique. A total of 1,275 nucleotides was examined in 5 sequences of DNA from Saccharomyces cerevisiae. Binding sites were non-random. The primary determinant of binding was the presence of a thymidine residue. Secondary determinants appeared to depend on the secondary structure of the DNA, with runs of thymidines providing unusually poor binding sites while TA and, especially, TATA providing the strongest binding sites. The 'TATA element' upstream of the URA3 gene from S. cerevisiae was the strongest binding site. The data indicate that ANB-spermine binding to DNA is a probe for DNA secondary structure and suggest a role for polyamines in regulating the structure of chromatin in vivo.

Azides

Behavior of a transitional tubulovesicular compartment at the cis side of the Golgi apparatus in in vivo fusion studies of mammalian cells.

We have investigated the behavior in in vivo cell fusion experiments of a transitional compartment lying between the endoplasmic reticulum and Golgi apparatus to determine if the compartment, as recognized by the antibody G1/93, might congregate in a similar manner to Golgi apparatus [W. C. Ho et al. (1990) Eur. J. Cell Biol. 52, 315-327]. The distributions of the transitional tubulovesicular compartment, endoplasmic reticulum, and Golgi apparatus in HeLa cells were assessed by immunofluorescent staining using mouse monoclonal antibody G1/93, mouse monoclonal antibody HP 24, and rabbit anti-galactosyltransferase, respectively. In agreement with previous results [W. C. Ho et al. (1990) Eur. J. Cell Biol. 52, 315-327], the Golgi apparatus was observed to congregate gradually over a 3- to 6-h period, forming a large, extended, central Golgi complex in uv-inactivated Sindbis virus-fused HeLa cells. Concomitant with this was a marked congregation of the transitional tubulovesicular compartment. Congregation of the tubulovesicular compartment was not affected by cycloheximide. The endoplasmic reticulum retained its web-like distribution throughout the syncytoplasm and rimmed the nuclear periphery. Treatment of HeLa cells with nocodazole prior to fusion followed by incubation of the syncytia in drug-containing media blocked congregation of the G1/93-positive compartment. With this long-term nocodazole treatment, Golgi apparatus was dispersed into scattered Golgi elements and the G1/93 distribution was endoplasmic reticulum-like. These results suggest that the transitional tubulovesicular compartment recognized by G1/93 is normally structured on microtubules and microtubule organizing centers and may be considered to be a subcompartment of a greater, perinuclear, Golgi complex.

Antibodies, Monoclonal

Hysteroscopic hydrotubation for treatment of tubal blockage.

In order to increase the effectiveness of hydrotubation, a plastic cannula was inserted into the fallopian tube under hysteroscopic control. Fifty women with tubal blockage confirmed by HSG were selected for treatment. Hydrotubation solution consisted of hydrocortisone (20 mg), gentamicin (160,000 IU) and procaine (80 mg) in 20 ml distilled water. Repeat HSG was performed. After three treatments, tubal patency (one or both tubes) was achieved in 20 women (40%), and seven pregnancies resulted within 1-18 months. Hysteroscopic hydrotubation may be an alternative treatment for tubal blockage.

Adult