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Biomedical subjects

H He

Publications and source records attributed to H He.

At least 19 recordsLinked to original sources

Analysis of cytokine regulators inducing interferon production by mouse uterine natural killer cells.

In mice and women, terminal differentiation of uterine natural killer (uNK) cells commences during endometrial decidualization. Both proliferation and interferon (IFN)-gamma are induced. Uterine NK cell precursors appear to home from secondary lymphoid organs to decidualizing uteri and localize mesometrially to the central decidua basalis, the site of maternal arterial modification at Gestation Days (gd) 9.5-10. In mice, genetic absence of uNK cells results in absence of pregnancy-induced spiral artery modification. Administration of IFN-gamma to uNK-negative pregnant females induces arterial modifications without fetal loss. In this study, we investigated the roles of cytokines, known in other tissues to differentiate and activate NK cells, in induction of IFN-gamma production in normal mouse implantation sites. Fecundity evaluation, implantation site morphometry, and IFN-gamma quantification in interleukin (IL)-12p40(0/0), IL-18(0/0), dual IL-12p40(0/0)/IL-18(0/0) and congenic strains revealed the importance of both IL-12 and IL-18 in the induction of spiral artery modification and IFN-gamma synthesis. Immediately after implantation, IL-18 was localized transiently to decidual cells, but by gd8, IL-18 was produced solely by uNK cells, suggesting that early uNK cells are activated by stroma and lymphocyte-derived signals maintain later uNK cell activation. Mesometrial tissue of C57Bl/6J mice was examined by reverse transcription polymerase chain reaction assay in virgin, early postimplantation, and midgestation females for expression of the heterodimeric cytokines IL-23 (composed of IL-12p40 and a novel alpha chain), IL-27 (composed of two IL-12-related chains) and IL-27R. No expression was detected in virgin uteri. The four genes were induced by gd6, and uNK cells isolated from midgestation transcribed IL-23alpha and IL-27R. This study advances the understanding of uNK cell activation during normal pregnancy.

Animals↗

Experimental lensless soft-X-ray imaging using iterative algorithms: phasing diffuse scattering.

Images of randomly placed two-dimensional arrays of gold balls have been reconstructed from their soft-X-ray transmission diffraction patterns. An iterative hybrid input-output (HiO) algorithm was used to solve the phase problem for the continuous distribution of diffuse X-ray scattering. Knowledge of the approximate size of the clusters was required. The images compare well with scanning electron microscope (SEM) images of the same sample. The use of micrometre-sized silicon nitride window supports is suggested, and absorption filters have been used to allow collection of low spatial frequencies often obscured by a beam stop. This method of phasing diffuse scattering may have application to scattering from individual inorganic nanostructures or single macromolecules.

Journal Article↗

Functional analysis of murine uterine natural killer cells genetically devoid of oestrogen receptors.

Uterine Natural Killer (uNK) cell differentiation in vivo requires oestrogen (E) priming prior to progesterone (P). Hybridomas between uNK precursor and SP2/0 cells express message for E receptor (ER)alpha but nor PR. However, mature, rodent and human uNK cells lack these receptors. To functionally assess requirements for uNK cell expression of ERalpha or ERbeta during precursor differentiation, marrow was transplanted from either ERalpha(o/o) (alphaERKO) or ERbeta(o/o) (betaERKO) mice into alymphoid RAG-2(o/o)/gammac(o/o) females. Recipients were mated and their implantation sites were examined by light microscopy, morphometry and ultrastructure. High numbers of uNK cells were established from each donor strain. Graft-derived uNK cells were similar in number and morphology to uNK cells of normal mice, suggesting that neither alpha- nor beta-ER is required for uNK precursor cell differentiation. Induction of spiral artery modification in the transplant recipients indicated that graft-derived uNK cells had functional properties. A novel technique for rapid isolation of highly purified uNK cells from normal mice using Dolichos biflorus agglutinin (DBA) lectin-conjugated magnetic beads was employed to obtain RNA. Expression of alpha- and beta-ER was absent by RT-PCR from NK cells isolated from the uterus, supporting the conclusions from the in vivo study.

Animals↗

Magnetic order and dynamics in an orbitally degenerate ferromagnetic insulator.

Neutron scattering was used to determine the spin structure and the magnon spectrum of the Mott-Hubbard insulator YTiO3. The magnetic structure is complex, comprising substantial G-type and A-type antiferromagnetic components in addition to the predominant ferromagnetic component. The magnon spectrum, on the other hand, is gapless and nearly isotropic. We show that these findings are inconsistent with the orbitally ordered states thus far proposed for YTiO3 and discuss general implications for a theoretical description of exchange interactions in orbitally degenerate systems.

Journal Article↗

Molecular cloning and nucleotide sequence of a new P450 gene, CYP319A1, from the cattle tick, Boophilus microplus.

We have isolated and sequenced a novel P450 gene (CYP319A1) from the cattle tick, Boophilus microplus. The CYP319A1 cDNA encodes a protein of 531 amino acids with an estimated molecular weight of 60.9k. It contains all highly conserved motifs characteristic of P450 enzymes. Comparison of deduced amino acid sequence with other CYP members shows that the CYP319A1 is more closely related to CYP4 family, but its overall identity to the CYP4 family is less than 40%. Therefore, it was assigned to a new P450 family by the P450 nomenclature committee. A pseudogene which shares high homology with the CYP319A1 was identified. Analysis of genomic sequence of the pseudogene indicated that the pseudogene contains two additional DNA inserts in the coding region, which disrupt the open reading frame. RT-PCR analysis showed that CYP319A1 is expressed in both susceptible and acaricide-resistant ticks.

Amino Acid Sequence↗

Magnetic resonant mode in the single-layer high-temperature superconductor Tl2Ba2CuO(6+delta).

An unusual spin excitation mode observed by neutron scattering has inspired numerous theoretical studies of the interplay between charged quasiparticles and collective spin excitations in the copper oxide high-temperature superconductors. The mode has, thus far, only been observed in materials with crystal structures consisting of copper oxide bilayers, and it is absent in the single-layer compound La(2-x)Sr(x)CuO(4+delta). Neutron-scattering data now show that the mode is present in Tl(2)Ba(2)CuO(6+delta), a single-layer compound with a superconducting transition temperature of approximately 90 kelvin, demonstrating that it is a generic feature of the copper oxide superconductors, independent of the layer sequence. This restricts the theoretical models for the origin of the resonant mode and its role in the mechanism of high-temperature superconductivity.

Journal Article↗

Crystallization and preliminary X-ray crystallographic analysis of native and selenomethionyl recombinant tabtoxin-resistance protein complexed with acetyl-coenzyme A.

Tabtoxin-resistance protein (TTR), an acetyltransferase from Pseudomonas syringae pv. tabaci, was overexpressed in Eschericha coli M15 and the TTR fusion protein complexed with acetyl-coenzyme A (AcCoA) was purified and crystallized. Diffraction data were collected to 3.0 A resolution in-house and the crystal was found to belong to space group P2(1), with unit-cell parameters a = 47.6, b = 66.6, c = 53.5 A, beta = 104.3 degrees. Furthermore, a selenomethionine (SeMet) TTR fusion protein derivative was overexpressed in the same expression system and its complex with AcCoA was purified in a reductive environment. The SeMet TTR derivative crystallized in two forms: the first was identical to that observed for native crystals and the second belonged to space group C2, with unit-cell parameters a = 101.7, b = 45.6, c = 84.2 A, beta = 105.8 degrees. Data from the P2(1) crystal form were collected in-house to 2.3 A resolution. Subsequently, three different wavelength data sets of the C2 crystal form to 1.55 A resolution were collected at the Advanced Photon Source at Argonne National Laboratory.

Acetyl Coenzyme A↗

[Comparison of (1)H MRS, PET and EEG in lateralization of temporal lobe epilepsy].

OBJECTIVE: To evaluate and compare the role of (1)H MRS, PET and EEG in lateralization diagnosis (LD) of temporal lobe epilepsy (TLE). METHODS: 15 patients with intractable TLE verified by operation and pathology were studied. Before unilateral anterior temporal lobectomy, (1)H MRS, PET and EEG were performed for the LD of TLE. The diagnostic value of (1)H MRS, PET and EEG was compared based on the pathology and follow-up results. RESULTS: With video scalp EEG monitoring, the focus was localized in one temporal lobe for 13 patients among which 5 had abnormal waves conducted to the contralateral lobe. Intracranial EEG was performed for other 2 patients because scalp EEG failed to make the LD. Ultimately, partial temporal lobectomy was performed in all patients based on LD by scalp EEG or intracranial EEG. The pathological findings of operation specimens showed various degree of hippocampal sclerosis. The surgery was proved to be effective by follow-ups of all 15 patients, with their conditions controlled in 12 cases and better in two. EEG succeeded to make correct LD in all cases. (1)H MRS made correct LD in 12 cases (80%) and incorrect LD in 1 case, and failed to made LD in 2 cases. Five bilateral abnormalities (33%) were found by MRS. FDG-PET made correct LD in 14 cases (93%) and incorrect LD in 1 case, and found two cases with bilateral abnormalities (13%). CONCLUSION: (1)H MRS, PET and EEG are all effective means in LD of TLE. Up to now, scalp EEG is the first choice and continuous video EEG monitoring has its own unique advantage. As a kind of simple, noninvasive and objective method, MRS can be used as a routine examination in conjunction with conventional MRI. Although PET is quite sensitive, it is expensive and has a number of limitations. So it can be used depending on the affordability of patients or diagnostic need. Combination of MRI, MRS, PET and scalp EEG may improve the reliability of the LD compared with use of any one of them alone. While the results of these methods are controversial, intracranial EEG is inevitable.

Adolescent↗

Synthesis and properties of fac-Re(dmbpy)(CO)3CHO (dmbpy = 4,4'-dimethyl-2,2'-bipyridine), a possible intermediate in reductions of CO2 catalyzed by fac-Re(dmbpy)(CO)3Cl.

Synthesis of fac-Re(dmbpy)(CO)3CHO 2 and its reactions with CO2 in DMF and DMSO have been conducted; 2 transfers hydride to CO2 to give Re(dmbpy)(CO)4+ OCHO- 5 which is rapidly transformed to fac-Re(dmbpy)(CO)3(OCHO) 3 in DMF, thus supporting the viability of 2 in photocatalytic reactions of fac-Re(dmbpy)(CO)3Cl with CO2.

Journal Article↗

Tricarbonyl[1,1',1"-ethylidynetris(pyrazole-kappaN2)]rhenium(I) bromide and tricarbonyl[methylidynetris(pyrazole-kappaN2)]rhenium(I) iodide ethanol hemisolvate.

The two title compounds, [Re(C(10)H(10)N(6))(CO)(3)]Br and [Re(C(11)H(12)N(6))(CO)(3)]I.0.5C(2)H(6)O, have slightly distorted octahedral geometries about the rhenium centers. The distortions result from the constraints of the eta(3)-coordinated tris(pyrazol-1-yl)methane ligands in each case which reduce the N-Re-N bond angles well below the preferred value of 90 degrees for facially disposed ligands at a six-coordinate metal center.

Journal Article↗

A tumor necrosis factor alpha- and interleukin 6-inducible protein that interacts with the small subunit of DNA polymerase delta and proliferating cell nuclear antigen.

A cDNA encoding a protein of 36 kDa, polymerase delta-interacting protein 1 (PDIP1), that interacts with the small subunit (p50) of DNA polymerase delta (pol delta) was identified in a two-hybrid screen of a HepG2 cDNA library by using p50 as bait. The interaction of PDIP1 with p50 was confirmed by pull-down assays, and a similar assay was used to demonstrate that PDIP1 interacts directly with the proliferating cell nuclear antigen (PCNA). PCNA and p50 bound to PDIP1 simultaneously, and PDIP1 stimulated pol delta activity in vitro in the presence, but not the absence, of PCNA, suggesting that PDIP1 also interacts functionally with both p50 and PCNA. Subcellular localization studies demonstrated that PDIP1 is a nuclear protein that colocalizes with PCNA at replication foci. A putative PCNA-binding motif was identified within the C terminus of PDIP1, and a synthetic peptide containing this PCNA-binding motif was shown to bind PCNA by far-Western analysis. Northern analysis demonstrated that PDIP1 mRNA is present in a wide variety of human tissues. PDIP1 was found to be highly homologous to a previously identified protein, B12 [Wolf, F. W., Marks, R. M., Sarma. V., Byers, M. G., Katz, R. W., Shows, T. B. & Dixit, V. M. (1992) J. Biol. Chem. 267, 1317-1326], one of the early response genes induced by tumor necrosis factor alpha. PDIP1 synthesis can also be induced by tumor necrosis factor alpha and by IL-6, cytokines essential for liver regeneration after loss of hepatic tissue. It is suggested that PDIP1 provides a link between cytokine activation and DNA replication in liver as well as in other tissues.

Amino Acid Sequence↗

Phosphorylation of mitochondrial elongation factor Tu in ischemic myocardium: basis for chloramphenicol-mediated cardioprotection.

The objective of this study was to identify the mitochondrial proteins that undergo changes in phosphorylation during global ischemia and reperfusion in the isolated rabbit heart. We also assessed whether the cardioprotective intervention of ischemic preconditioning affected mitochondrial protein phosphorylation. We established a reconstituted system using isolated mitochondria and cytosol from control or ischemic hearts. We found that phosphorylation of a 46-kDa protein on a serine residue was increased in ischemia and that phosphorylation was reduced in control or preconditioned hearts. Using 2D gel electrophoresis and mass spectrometry, we have identified the 46-kDa protein as mitochondrial translational elongation factor Tu (EF-Tu(mt)). These data reveal that ischemia and preconditioning modulate the phosphorylation of EF-Tu(mt) and suggest that the mitochondrial protein synthesis machinery may be regulated by phosphorylation. Phosphorylation of mitochondrial EF-Tu has not been previously described; however, in prokaryotes, EF-Tu phosphorylation inhibits protein translation. We hypothesized that phosphorylation of mitochondrial EF-Tu would inhibit mitochondrial protein translation and attempted to reproduce the effect with inhibition of mitochondrial protein synthesis by chloramphenicol. We found that chloramphenicol pretreatment significantly reduced infarct size, suggesting that mitochondrial protein synthesis is one determinant of myocardial injury during ischemia and reperfusion.

Adenosine Triphosphate↗

Administration of pentoxifylline during allergen sensitization dissociates pulmonary allergic inflammation from airway hyperresponsiveness.

Asthma, a chronic inflammatory disease characterized by intermittent, reversible airflow obstruction and airway hyperresponsiveness (AHR), is classically characterized by an excess of Th2 cytokines (IL-13, IL-4) and depletion of Th1 cytokines (IFN-gamma, IL-12). Recent studies indicating an important role for Th1 immunity in the development of AHR with allergic inflammation suggest that Th1/Th2 balance may be important in determining the association of AHR with allergic inflammation. We hypothesized that administration of pentoxifylline (PTX), a phosphodiesterase inhibitor known to inhibit Th1 cytokine production, during allergen (OVA) sensitization and challenge would lead to attenuation of AHR in a murine model of allergic pulmonary inflammation. We found that PTX treatment led to attenuation of AHR when administered at the time of allergen sensitization without affecting other hallmarks of pulmonary allergic inflammation. Attenuation of AHR with PTX treatment was found in the presence of elevated bronchoalveolar lavage fluid levels of the Th2 cytokine IL-13 and decreased levels of the Th1 cytokine IFN-gamma. PTX treatment during allergen sensitization leads to a divergence of AHR and pulmonary inflammation following allergen challenge.

Aerosols↗

[Tissue tropism of novel nonenveloped DNA virus in experimentally infected rhesus monkey].

OBJECTIVE: To investigate the tissue tropism of transfusion transmitted virus (TTV), a novel nonenveloped DNA virus, in experimentally infected Rhesus monkey. METHODS: Filtrate of feces of one patient with TTV infection was fed into the stomachs of ten Rhesus monkeys. Five (?) monkeys were killed during viremia and their tissues were collected to be examined for the presence of replicative virus DNA by in situ hybridization, dot blot hybridization with antisense probe and hybridization/nuclease protection assay. RESULTS: The virus contents (density of electrophoretic band) in the liver, PBMC, and serum of three infected monkeys 50.1%, 31.0%, and 18.9% respectively; the relative proportion was 2.65;1.64;1.00. TTV was detected in liver, spleen, stomach, small intestine and its lymph nodes, PBMC, colon, and serum; however, viral plus-chain, a putative replicative intermediate, was found only in liver, spleen, small intestine, and lymphoid cells. CONCLUSION: Transfusion transmitted virus is hepato-, spleno-, and lymphoid cell-tropic, and may has multiple routes of transmission.

Animals↗

Saccharomyces cerevisiae protein Pci8p and human protein eIF3e/Int-6 interact with the eIF3 core complex by binding to cognate eIF3b subunits.

Mammalian, plant, and Schizosaccharomyces pombe eukaryotic initiation factor-3 (eIF3) contains a protein homologous to the product of int-6 (eIF3e), a frequent integration site of mouse mammary tumor viruses. By contrast, Saccharomyces cerevisiae does not encode a protein closely related to eIF3e/Int-6. Here, we characterize a novel S. cerevisiae protein (Pci8p, Yil071cp) that contains a PCI (proteasome-COP9 signalosome-eIF3) domain conserved in eIF3e/Int-6. We show that both Pci8p and human eIF3e/Int-6 expressed in budding yeast interact with the yeast eIF3 complex in vivo and in vitro by binding to a discrete segment of its eIF3b subunit Prt1p and that human eIF3e/Int-6 interacts with the human eIF3b segment homologous to the Pci8p-binding site of yeast Prt1p. These results refine our understanding of subunit interactions in the eIF3 complex and suggest structural similarity between human eIF3e/Int-6 and yeast Pci8p. However, deletion of PCI8 had no discernible effect on cell growth or translation initiation as judged by polysome analysis, suggesting that Pci8p is not required for the essential function of eIF3 in translation initiation. Motivated by the involvement of Int-6 in transcriptional control, we investigated the effects of deleting PCI8 on the total mRNA expression profile by oligonucleotide microarray analysis and found reduced mRNA levels for a subset of heat shock proteins in the pci8Delta mutant. We discuss possible dual functions of Pci8p and Int-6 in transcriptional and translational control.

Binding Sites↗

Bid is cleaved by calpain to an active fragment in vitro and during myocardial ischemia/reperfusion.

Reperfusion after myocardial ischemia is associated with a rapid influx of calcium, leading to activation of various enzymes including calpain. Isolated perfused adult rabbit hearts subjected to global ischemia and reperfusion were studied. Calpain or a calpain-like activity was activated within 15 min after reperfusion, and preconditioning suppressed calpain activation. In contrast, caspase activation was not detected although cytochrome c was released after ischemia and reperfusion. The pro-apoptotic BH3-only Bcl-2 family member, Bid, was cleaved during ischemia/reperfusion in the adult rabbit heart. Recombinant Bid was cleaved by calpain to a fragment that was able to mediate cytochrome c release. The calpain cleavage site was mapped to a region within Bid that is extremely susceptible to proteolysis. These findings suggest that there is cross-talk between apoptotic and necrotic pathways in myocardial ischemia/reperfusion injury.

Amino Acid Sequence↗