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PubMed2005

Two-photon spectroscopy of core excitons in barium fluoride using synchrotron radiation and laser light.

We have conducted two-photon spectroscopy of core excitons in BaF(2). Synchrotron radiation and laser light were used for 5p core-electron excitation and Auger-free luminescence was detected as the signal. Two-photon excitation enables access to f and p orbitals that cannot be reached by one-photon excitation of electrons in p orbitals. It has been found that the spin-orbit splittings of 4f and 6p states of the Ba ion in BaF(2) are 0.7 +/- 0.1 and 1.4 +/- 0.1 eV, respectively.

Journal Article↗

PubMed2002

The ordered hydrocerussite-type structure of (PbCO3)2.BaF2.

Single crystals of synthetic (PbCO(3))(2).BaF(2), bis[lead(II) carbonate] barium difluoride, have been grown under hydrothermal conditions. The compound crystallizes in the ordered hydrocerussite [(PbCO(3))(2).Pb(OH)(2)] structure and can be derived from a close packing of the metal atoms with a stacking sequence of [c(h)(2)](3) along the c axis. O and F atoms are situated approximately in the tetrahedral voids, resulting in the formation of a slightly distorted [BaF(6)O(6)] icosahedron and a [1+6+3]-coordinate Pb atom, with one short bond to F, six bonds to O and three longer bonds to additional O atoms. The carbonate group deviates only slightly from the geometry of an equilateral triangle.

Journal Article↗

PubMed1992

Ozone depletion and its effects on human populations.

Concern about the ozone-depleting potential of man-made chemicals has led the United Nations to control their usage. Under the present arrangements, fully halogenated chlorofluorocarbons, halons and carbon tetrachloride will be phased out by the year 2000, and methylchloroform by the year 2005. Even so, atmospheric chlorine is expected to rise to seven times its natural level, so further ozone losses and greater ultraviolet B (UVB) exposure can be expected over heavily populated areas of the globe. Calculating changes in skin cancer incidence is a two-stage process which must take account of the increase in biologically effective UVB that results from an ozone loss of 1% (optical amplification factor, OAF) and the percentage increase in skin cancer incidence that results from a 1% increase in annual UV dose (biological amplification factor, BAF). Epidemiological data provide a BAF value of approximately 1.7 for basal cell carcinoma and 3.0 for squamous cell carcinoma. The absorption spectrum of ozone and the increased carcinogenic impact of UV radiation around 300 nm results in a value for OAF of approximately 1.6%; thus a 10% loss of ozone, if sufficiently sustained, would eventually increase the incidence of basal and squamous cell carcinomas by almost 30% and 50%, respectively. Dermatologists, therefore, need to look carefully at the environment in order to safeguard the health of future generations.

Air Pollution↗

PubMed1994

Pharmacokinetic-pharmacodynamic modeling between pinacidil or pinacidil-N-oxide plasma levels and systemic and regional hemodynamic effects in healthy volunteers.

Pinacidil (P) lowers blood pressure through peripheral vasodilation, but also induces dose-dependent side-effects. In a previous placebo-controlled, randomized, double-blind and crossover study, performed in six healthy male volunteers, we investigated the systemic and regional hemodynamic effects of a single oral administration of 25 mg of P (sustained-release form) and measured the plasma concentrations of P and of its active metabolite, pinacidil-N-oxide (PO). In the present study, our goal has been to investigate the relationships between P and/or PO plasma concentrations and P administration effects on systolic, diastolic and mean arterial pressures (SAP, DAP, MAP), heart rate (HR), cardiac output (CO), total peripheral resistance (TPR), brachial and carotid arteries' diameters (BAD, CAD), flows (BAF, CAF) and vascular resistances (BVR, CVR) which were assessed before and at different time intervals after drug intake. Concentration-effect relationships were investigated using both linear and log-linear multiple regression models with P, PO or both P and PO as independent variables (six models). Significant linear relationships were observed between P and/or PO and SAP, DAP, MAP, TPR, BAD, BAF, BVR, CAD and CVR. For example, TPR (dynes.s/cm5) = 1308-3.031 x P (ng/ml), R = 0.57, P = 0.0037; BVR (mmHg.s/ml) = 58-0.261 x P (ng/ml), R = 0.56, P = 0.0042. Almost similar R values were obtained using P, PO, or both P and PO. The use of log-linear models did not improve the fittings.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1996

Differential interaction with endocytic and exocytic pathways distinguish parasitophorous vacuoles of Coxiella burnetii and Chlamydia trachomatis.

Coxiella burnetii and Chlamydia trachomatis are bacterial obligate intracellular parasites that occupy distinct vacuolar niches within eucaryotic host cells. We have employed immunofluorescence, cytochemistry, fluorescent vital stains, and fluid-phase markers in conjunction with electron, confocal, and conventional microscopy to characterize the vacuolar environments of these pathogens. The acidic nature of the C. burnetii-containing vacuole was confirmed by its acquisition of the acidotropic base acridine orange (AO). The presence of the vacuolar-type (H+) ATPase (V-ATPase) within the Coxiella vacuolar membrane was demonstrated by indirect immunofluorescence, and growth of C. burnetii was inhibited by bafilomycin A1 (Baf A), a specific inhibitor of the V-ATPase. In contrast, AO did not accumulate in C. trachomatis inclusions nor was the V-ATPase found in the inclusion membrane. Moreover, chlamydial growth was not inhibited by Baf A or the lysosomotropic amines methylamine, ammonium chloride, and chloroquine. Vacuoles harboring C. burnetii incorporated the fluorescent fluid- phase markers, fluorescein isothiocyanate-dextran (FITC-dex) and Lucifer yellow (LY), indicating trafficking between that vacuole and the endocytic pathway. Neither FITC-dex nor LY was sequestered by chlamydial inclusions. The late endosomal-prelysosomal marker cation-independent mannose 6-phosphate receptor was not detectable in the vacuolar membranes encompassing either parasite. However, the lysosomal enzymes acid phosphatase and cathepsin D and the lysosomal glycoproteins LAMP-1 and LAMP-2 localized to the C. burnetii vacuole but not the chlamydial vacuole. Interaction of C. trachomatis inclusions with the Golgi-derived vesicles was demonstrated by the transport of sphingomyelin, endogenously synthesized from C6-NBD-ceramide, to the chlamydial inclusion and incorporation into the bacterial cell wall. Similar trafficking of C-NBD-ceramide was not evident in C. burnetii-infected cells. Collectively, the data indicate that C. trachomatis replicates within a nonacidified vacuole that is disconnected from endosome-lysosome trafficking but may receive lipid from exocytic vesicles derived from the trans-Golgi network. These observations are in sharp contrast to those for C. burnetii, which by all criteria resides in a typical phagolysosome.

Animals↗

PubMed2006

Wild-type levels of human immunodeficiency virus type 1 infectivity in the absence of cellular emerin protein.

Preintegration complexes (PICs) mediate retroviral integration, and recent results indicate an important role for the inner nuclear membrane protein emerin in orienting human immunodeficiency virus type 1 (HIV-1) PICs to chromatin for integration. Two other host cell proteins, the barrier-to-autointegration factor (BAF) and lamina-associated polypeptide 2alpha (LAP2alpha), seemed to play a similar preintegrative role for Moloney murine leukemia virus (MMLV) in addition to HIV-1. In contrast, we determined efficient HIV-1 and MMLV infection of HeLa-P4 cells following potent down-regulation of emerin, BAF, or LAP2alpha protein by using short interfering RNA. Mouse embryo fibroblasts ablated for emerin protein through gene knockout support the same level of HIV-1 infection as cells derived from wild-type littermate control animals. As the expression of human emerin in mouse knockout cells fails to affect the level of infectivity achieved in its absence, we conclude that HIV-1 efficiently infects cells in the absence of emerin protein and, by extension, that emerin is not a universally important regulator of HIV-1 infectivity.

Amino Acid Sequence↗

PubMed2001

Adeno-associated virus type 2-mediated gene transfer: altered endocytic processing enhances transduction efficiency in murine fibroblasts.

Adeno-associated virus type 2 (AAV) is a single-stranded-DNA-containing, nonpathogenic human parvovirus that is currently in use as a vector for human gene therapy. However, the transduction efficiency of AAV vectors in different cell and tissue types varies widely. In addition to the lack of expression of the viral receptor and coreceptors and the rate-limiting viral second-strand DNA synthesis, which have been identified as obstacles to AAV-mediated transduction, we have recently demonstrated that impaired intracellular trafficking of AAV inhibits high-efficiency transduction of the murine fibroblast cell line, NIH 3T3 (J. Hansen, K. Qing, H. J. Kwon, C. Mah, and A. Srivastava, J. Virol. 74:992-996, 2000). In this report, we document that escape of AAV from the endocytic pathway in NIH 3T3 cells is not limited but processing within endosomes is impaired compared with that observed in the highly permissive human cell line 293. While virions were found in both early and late endosomes or lysosomes of infected 293 cells, they were localized predominantly to the early endosomes in NIH 3T3 cells. Moreover, treatment of cells with bafilomycin A1 (Baf), an inhibitor of the vacuolar H(+)-ATPase and therefore of endosomal-lysosomal acidification, decreased the transduction of 293 cells with a concomitant decrease in nuclear trafficking of AAV but had no effect on NIH 3T3 cells. However, after exposure of NIH 3T3 cells to hydroxyurea (HU), a compound known to increase AAV-mediated transduction in general, virions were detected in late endosomes and lysosomes, and these cells became sensitive to Baf-mediated inhibition of transduction. Thus, HU treatment overcomes defective endocytic processing of AAV in murine fibroblasts. These studies provide insights into the underlying mechanisms of intracellular trafficking of AAV in different cell types, which has implications in the optimal use of AAV as vectors in human gene therapy.

3T3 Cells↗

PubMed1998

A critical role for cyclin C in promotion of the hematopoietic cell cycle by cooperation with c-Myc.

Cyclin C, a putative G1 cyclin, was originally isolated through its ability to complement a Saccharomyces cerevisiae strain lacking the G1 cyclin gene CLN1-3. Unlike cyclins D1 and E, the other two G1 cyclins obtained by the same approach and subsequently shown to play important roles during the G1/S transition, there is thus far no evidence to support the hypothesis that cyclin C is indeed critical for the promotion of cell cycle progression. In BAF-B03 cells, an interleukin 3 (IL-3)-dependent murine pro-B-cell line, cyclin C gene mRNA was induced at the G1/S phase upon IL-3 stimulation and reached a maximal level in the S phase. Enforced expression of exogenous cyclin C in this cell line failed to alter its growth properties. In the present study, we examined whether cyclin C is capable of cooperating with the cytokine-responsive immediate-early gene products c-Myc and c-Fos in the promotion of cell proliferation. We found that cyclin C is able to cooperate functionally with c-Myc, but not c-Fos, to induce both BAF-B03 cell proliferation in a cytokine-independent fashion and the formation of cell clusters. Furthermore, cyclin C was primarily responsible for the induction of cdc2 gene expression. Our data define a novel role for cyclin C in the regulation of both the G1/S and G2/M phases of the cell cycle, and this effect appears to be independent of the activity of CDK8 in the control of transcription.

Animals↗

PubMed2001

Nuclear actin and actin-related proteins in chromatin remodeling.

The existence and function of actin in the nucleus has been hotly debated for forty years. Recently, beta-actin was found to be a component of mammalian SWI/SNF-like BAF chromatin remodeling complexes and still more recently other SWI/SNF-related chromatin remodeling complexes in yeast, flies, and man. Although the function of actin in these chromatin remodeling complexes is only starting to be explored, the fact that actin is one of the most regulated proteins in the cell suggests that control of nuclear actin may be a critical regulatory point in the control of chromatin remodeling. Actin rapidly shuttles between the nucleus and the cytoplasm offering additional sites and modes of regulation. In addition, actin-related proteins (Arps) are also components of these chromatin remodeling complexes and have been implicated in transcriptional control in yeast. The observation that the BAF chromatin remodeling complex in which actin was originally identified, is also a human tumor suppressor complex necessary for the actions of the retinoblastoma protein indicates that the study of nuclear actin is likely to contribute to understanding cell growth control.

Actins↗

PubMed1981

The effect of epidermal growth factor (EGF) on the differentiation of the rough endoplasmic reticulum in fetal mouse small intestine in organ culture.

The differentiation of the rough endoplasmic reticulum (RER) of mouse duodenal absorptive cells located at the tip of the villi at 17 days of gestation was compared to that of absorptive cells in duodenal explants of 15-day-old mouse fetuses cultured for 72 hr 1) with Trowell T8 medium (without insulin) alone or supplemented 2) with epidermal growth factor (EGF; 100 ng/ml) or 3) with 25% bovine amniotic fluid (BAF). Glucose-6-phosphatase activity (G6Pase) was localized cytochemically to ensure a better identification of the RER. The intersections of a double lattice falling over and outside the RER were counted and the percentage of intersections over the RER was estimated. With this method, the extent of the RER is not statistically different when the absorptive cells in utero are compared to those of explants cultured with EGF. However, the extent of the RER in the absorptive cells cultured with Trowell T8 medium alone or supplemented with BAF is 50% lower than in the former two groups. It is concluded that EGF promotes the maturation of duodenal absorptive cells in organ culture.

Animals↗

PubMed2001

Perverted responses of the human granulocyte-macrophage colony-stimulating factor receptor in mouse cell lines due to cross-species beta-subunit association.

Transfected murine cell lines are commonly used to study the function of many human cytokine or receptor mutants. This study reports the inappropriate activation of the human granulocyte-macrophage colony-stimulating factor (hGM-CSF) receptor by the human GM-CSF antagonist, E21R, when the human receptor is introduced into the murine cell line BaF-B03. E21R-induced proliferation of the BaF-B03 cells is dependent on transfection with both hGM-CSF receptor alpha and beta(c) subunits. Studies on the underlying mechanism revealed constitutive association between human and mouse beta(c) and GM-CSF receptor-alpha, tyrosine phosphorylation of mouse and human beta(c), and association of phosphorylated mouse beta(c) into an activated human GM-CSF receptor complex in response to E21R and GM-CSF. This interspecies receptor cross-talk of receptor signaling subunits may produce misleading results and emphasizes the need to use cell lines devoid of the cognate endogenous receptors for functional analysis of ligand and receptor mutants.

Animals↗

PubMed2005

LEM2 is a novel MAN1-related inner nuclear membrane protein associated with A-type lamins.

The LEM (lamina-associated polypeptide-emerin-MAN1) domain is a motif shared by a group of lamin-interacting proteins in the inner nuclear membrane (INM) and in the nucleoplasm. The LEM domain mediates binding to a DNA-crosslinking protein, barrier-to-autointegration factor (BAF). We describe a novel, ubiquitously expressed LEM domain protein, LEM2, which is structurally related to MAN1. LEM2 contains an N-terminal LEM motif, two predicted transmembrane domains and a MAN1-Src1p C-terminal (MSC) domain highly homologous to MAN1, but lacks the MAN1-specific C-terminal RNA-recognition motif. Immunofluorescence microscopy of digitonin-treated cells and subcellular fractionation identified LEM2 as a lamina-associated protein residing in the INM. LEM2 binds to the lamin C tail in vitro. Targeting of LEM2 to the nuclear envelope requires A-type lamins and is mediated by the N-terminal and transmembrane domains. Highly overexpressed LEM2 accumulates in patches at the nuclear envelope and forms membrane bridges between nuclei of adjacent cells. LEM2 structures recruit A-type lamins, emerin, MAN1 and BAF, whereas lamin B and lamin B receptor are excluded. Our data identify LEM2 as a novel A-type-lamin-associated INM protein involved in nuclear structure organization.

Animals↗

PubMed1990

Products of the fos and jun proto-oncogenes bind cooperatively to the AP1 DNA recognition sequence.

The products of the proto-oncogenes c-fos and c-jun form a tight protein complex that is a major component of the transcription factor AP1. To analyze the role of fos in the binding of this complex to the AP1 DNA recognition sequence and the mechanism of interaction in further detail, we have expressed a fos protein in E. coli using an expression vector containing the temperature-inducible lambda PL promoter and a synthetic translational start codon. The fos protein encoded by this construct (termed Baf) was enriched by biochemical purification techniques and was found to form a specific complex with c-jun obtained by in vitro transcription/translation. As shown in gel retardation assays, the baf/jun complex binds to the AP1 DNA recognition sequence with high affinity, while no significant binding was observed with either of the individual protein components, indicating cooperative DNA binding of the two proteins. The fact that the bacterial baf protein does not undergo glycosylation indicates that the post-translational modification of eukaryotic c-fos with N-acetylglucosamine is not required for the formation of a stable fos/jun/DNA complex.

Binding Sites↗

PubMed2005

Metabolic activity: a novel indicator of neuronal survival in the murine dopaminergic cell line CAD.

Apoptosis is implicated in many neurodegenerative diseases, including Parkinson's disease (PD). Neuroprotective strategies targeting apoptosis need to preserve functional integrity of the saved cells to be effective. The aim of the present study was to evaluate a novel approach for analyzing neuronal function that monitors cellular metabolic responses to receptor activation using the microphysiometer. N-Acetyl-sphingosine (C2-ceramide) induced cell death of the neuronal cell line, Cath.a-differentiated (CAD) cells, which resemble catecholaminergic cells of the CNS, and provide a useful in vitro model for the cells affected in PD. C2-ceramide also suppressed the metabolic response of CAD cells to muscarinic receptor activation. Pretreatment with the caspase inhibitor Boc-Asp-(OMe)-fluoromethylketone (BAF) plus neurotrophin-3 (NT-3) reduced C2- ceramide-induced CAD cell death, delaying cell death more effectively than either agent alone; and, most significantly, BAF and NT-3 enabled the cells remaining 24 h after toxin treatment to generate a normal metabolic response to the muscarinic agonist carbachol. On the basis of these results, we suggest that measuring metabolic responses to receptor activation is a useful method for following neuronal viability after toxin treatment and that the combination of caspase inhibitors and neurotrophic factors might be a plausible strategy for improving neuronal survival, with critical preservation of metabolic function.

Amino Acid Chloromethyl Ketones↗

PubMed2005

Bioaccumulation of metals in Eisenia fetida after exposure to a metal-loaded bauxsol reagent.

The present study investigated the acute toxic effects of a metal-loaded Bauxsol reagent (MLBR), containing more than 6,450 mg kg(-1) of bound metal, on the earthworm Eisenia fetida. Bauxsol is manufactured by Virotec Global Solutions of Gold Coast, Queensland, Australia. The E. fetida were exposed to 0, 10, 20, 40, 60, and 80% treatments of MLBR plus cow manure for 28 d. Good motility and no mortality of E. fetida were found in all treatments after 28 d of exposure. The greatest bioaccumulation of metals by E. fetida occurred in the 20 and 40% MLBR treatments. The bioaccumulation of metals in the worms and bioaccumulation factors (BAFs) were less than the reported toxicity thresholds to cause mortality and less than the reported BAF for moderately contaminated soils, indicating that metals bound to Bauxsol reagents are mostly nonbioavailable. Analyses of the 20% MLBR treatment at 28 d using a sequential extraction procedure showed that more than 95% of the metals are bound within the Fe/Mn oxide fractions. However, changes occurred in metal fractionation after exposure to E. fetida for Cd/Cr, and Fe/Mn. The data also showed that the exchangeable (1 M MgCl2) and the toxicity characteristic leaching procedure extractant are useful as indicators of metal bioavailability from MLBR to E. fetida.

Aluminum↗

PubMed2006

Oral glucosamine for 6 weeks at standard doses does not cause or worsen insulin resistance or endothelial dysfunction in lean or obese subjects.

Glucosamine is a popular nutritional supplement used to treat osteoarthritis. Intravenous administration of glucosamine causes insulin resistance and endothelial dysfunction. However, rigorous clinical studies evaluating the safety of oral glucosamine with respect to metabolic and cardiovascular pathophysiology are lacking. Therefore, we conducted a randomized, placebo-controlled, double-blind, crossover trial of oral glucosamine at standard doses (500 mg p.o. t.i.d.) in lean (n = 20) and obese (n = 20) subjects. Glucosamine or placebo treatment for 6 weeks was followed by a 1-week washout and crossover to the other arm. At baseline, and after each treatment period, insulin sensitivity was assessed by hyperinsulinemic-isoglycemic glucose clamp (SI(Clamp)) and endothelial function evaluated by brachial artery blood flow (BAF; Doppler ultrasound) and forearm skeletal muscle microvascular recruitment (ultrasound with microbubble contrast) before and during steady-state hyperinsulinemia. Plasma glucosamine pharmacokinetics after oral dosing were determined in each subject using a high-performance liquid chromatography method. As expected, at baseline, obese subjects had insulin resistance and endothelial dysfunction when compared with lean subjects (SI(Clamp) [median {25th-75th percentile}] = 4.3 [2.9-5.3] vs. 7.3 [5.7-11.3], P < 0.0001; insulin-stimulated changes in BAF [% over basal] = 12 [-6 to 84] vs. 39 [2-108], P < 0.04). When compared with placebo, glucosamine did not cause insulin resistance or endothelial dysfunction in lean subjects or significantly worsen these findings in obese subjects. The half-life of plasma glucosamine after oral dosing was approximately 150 min, with no significant changes in steady-state glucosamine levels detectable after 6 weeks of therapy. We conclude that oral glucosamine at standard doses for 6 weeks does not cause or significantly worsen insulin resistance or endothelial dysfunction in lean or obese subjects.

Administration, Oral↗

PubMed2001

Aneurysms of the fenestrated basilar artery treated with Gulielmi Detachabe Coils: case report.

Multiple aneurysms at the site of a basilar artery fenestration (BAF) are extremely rare. We report a case of BAF in association with two aneurysms successfully treated with Guglielmi Detachable Coils (GDCs). A 42-year-old male presented with subarachnoid hemorrhage and consciousness change. Angiograms of the vertebrobasilar artery demonstrated fenestration of the proximal basilar artery associated with two aneurysms. Embolization of aneurysms was done by using GDCs. Aneurysms were almost completely obliterated with preservation of the flow and lumen of the vertebrobasilar system. The patient remains asymptomatic at clinical follow-up of 18 months.

Adult↗

PubMed2001

Advanced biological unit processes for domestic water recycling.

The potential of advanced biological unit operations for the recycling of grey and black waters has been evaluated. The membrane bioreactor (MBR) demonstrated the greatest efficacy towards water recycling in terms of all the quality determinants. Both the biologically aerated filter (BAF) and the MBR were able to effectively treat the organic and physical pollutants in all the types of wastewater tested. The main difference was observed in terms of the microbiological quality, measured as total coliforms. The open bed structure of the BAF enabled passage of coliforms whereas the complete barrier of the MBR produced a non detectable level in the effluent. The MBR process complied with commonly adopted water recycling quality standards for the all determinants during the grey water trials and failed only in terms of total coliform counts once black water had been introduced into the feed. The MBR was seen as a particularly suitable advanced biological process as it was very effective at stabilising out the considerable load variations encountered during the trial.

Bioreactors↗