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Pflugers Arch. 2000 Oct;440(6):819-27.
Transport of magnesium by two isoforms of the Na+-Ca2+ exchanger expressed in CCL39 fibroblasts.

Tashiro M, Konishi M, Iwamoto T, Shigekawa M, Kurihara S.

Department of Physiology, The Jikei University School of Medicine, Tokyo, Japan.

Cytoplasmic concentrations of Ca2+ ([Ca2+]i) and Mg2+ ([Mg2+]i) were measured with fluorescent indicators in CCL39 cells, a cell line established from Chinese hamster lung fibroblasts, transfected with complementary deoxyribonucleic acid (cDNA) of the Na+-Ca2+ exchanger isolated either from canine heart (NCX1) or from rat brain (NCX3). Raising extracellular [Mg2+] to 10 mM increased Mg2+ influx and the resultant change in [Mg2+]i (delta[Mg2+]i) was monitored with furaptra under Ca2+-free conditions. In control (vector-transfected) cells, delta[Mg2+]i at 45 min was similar with or without extracellular Na+ (130 mM or 0 mM) and when [Na+]i was raised by 1 mM ouabain treatment. delta[Mg2+]i in NCX1-transfected cells was attenuated significantly in the presence of 130 mM Na+, but became comparable to (or slightly larger than) that in control cells on either removal of extracellular Na+ or treatment with 1 mM ouabain. Cells expressing NCX3 showed an intermediate dependence of delta[Mg2+]i on Na+, probably reflecting a lower degree of expression of the exchanger protein. Extracellular Na+-dependent changes in [Ca2+]i (measured with fura-2 in the presence of extracellular Ca2+ and 10 microM ionomycin, a Ca2+ ionophore) were minimal in control cells, marked in the NCX1-transfected cells and intermediate in the NCX3-transfected cells. These results suggest that the Na+-Ca2+ exchanger (either NCX1 or NCX3) can transport Mg2+ and may play a role in the extrusion of magnesium from cells.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11041546&dopt=Abstract

J Comp Pathol. 2000 Nov;123(4):302-5.
Distribution of porcine parvovirus in porcine circovirus 2-infected pigs with postweaning multisystemic wasting syndrome as shown by in-situ hybridization.

Choi C, Chae C.

Department of Veterinary Pathology, College of Veterinary Medicine, Seoul National University, Suwon 441-744, Kyounggi-Do, Republic of Korea.

In-situ hybridization with a nonradioactive digoxigenin-labelled probe was used to study the distribution of porcine parvovirus (PPV) in formalin-fixed paraffin wax-embedded tissues from 10 porcine circovirus 2 (PCV2)-infected weaned pigs with naturally occurring postweaning multisystemic wasting syndrome (PMWS). A 226 base pair DNA fragment from a VP2 structural gene was generated by polymerase chain reaction (PCR) and used as a probe. Lymph node, spleen, thymus and tonsil were positive by PCR, demonstrating the presence of PPV DNA in the tissue samples from four of 10 pigs tested. PPV nucleic acid was also detected consistently in lymph node, spleen, thymus and tonsil by in-situ hybridization. Detection of PPV DNA from PCV2-infected pigs with PMWS suggests that PPV also plays a role in the pathogenesis of PMWS. 2000 Harcourt Publishers Ltd.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11042001&dopt=Abstract

Eur J Pharm Sci. 2000 Sep;11(3):199-205.
Biodegradable pH-sensitive surfactants (BPS) in liposome-mediated nucleic acid cellular uptake and distribution.

Liang E, Rosenblatt MN, Ajmani PS, Hughes JA.

Department of Pharmaceutics, College of Pharmacy, University of Florida, P.O. Box 100494, Gainesville, FL 32610, USA.

The impact of biodegradable pH-sensitive surfactant (BPS)-liposomes on nucleic acid, i.e., oligonucleotide and plasmid DNA, cellular delivery was examined. Fluorescein-labeled nucleic acids complexed with 1,2-dioleoyl-3-trimethylammonium propane cationic liposomes and BPS at a charge ratio (+/-) of 10 were incubated in CV-1 cells and analyzed by flow cytometry. The fluorescence intensity of oligonucleotides but not plasmid DNA complexed with BPS-liposomes was higher than those complexed with BPS-free liposomes at early time points. However, when cells were fixed to equalize the intracellular pH since fluorescein, a pH-sensitive fluorophore, has higher fluorescence intensity in alkaline pH than acidic, no difference in intensity was observed. This indicated the incorporation of BPS in liposomes did not increase oligonucleotide cellular uptake over control liposomes, but redistributed oligonucleotides into a more basic environment, e.g., cytoplasm. An explanation consistent with the presented data is the formation of small transient membrane defects within the endosomal membrane as presented previously [Liang, E., Hughes, J.A., 1998a. Membrane fusion and rupture in liposomes: effect of biodegradable pH-sensitive surfactants. J. Membr. Biol. 166, 37-49.]. The above findings suggested that BPS may be effective agents of disrupting one of the major barriers, endosomal membrane, to enhance nucleic acid cellular transport.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11042225&dopt=Abstract

Neuroreport. 2002 Nov 15;13(16):2035-9.
Neuronal RNA oxidation is a prominent feature of dementia with Lewy bodies.

Nunomura A, Chiba S, Kosaka K, Takeda A, Castellani RJ, Smith MA, Perry G.

Department of Psychiatry and Neurology, Asahikawa Medical College, Higashi 2-1-1-1, Midorigaoka, Asahikawa 078-8510, Japan. nunsahika-med.ac.jp

An approach was used to identify the oxidized nucleoside, 8-hydroxyguanosine in brains of dementia with Lewy bodies. Neurons with marked immunoreaction of 8-hydroxyguanosine in the cytoplasm were widely distributed in the hippocampal region and temporal neocortex. Relative intensity measurements of neuronal 8-hydroxyguanosine immunoreactivity showed that there was a significant increase in nucleic acid oxidation in dementia with Lewy bodies compared with controls. Treatment with nuclease (DNase or RNase) before the immunostaining demonstrated that RNA was a major site of nucleic acid oxidation. Together with the previously reported RNA oxidation in vulnerable neurons in Alzheimer and Parkinson diseases, neuronal RNA oxidation in dementia with Lewy bodies might represent one of the fundamental abnormalities in age-associated neurodegenerative diseases.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12438921&dopt=Abstract

J Chromatogr A. 2000 Sep 29;893(1):23-35.
Preparation and evaluation of packed capillary columns for the separation of nucleic acids by ion-pair reversed-phase high-performance liquid chromatography.

Oberacher H, Krajete A, Parson W, Huber CG.

Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University, Innsbruck. Austria.

Oligonucleotides and double stranded DNA fragments were separated in 200 microm I.D. capillary columns packed with micropellicular, octadecylated, 2.1 microm poly(styrene-divinylbenzene) particles by ion-pair reversed-phase high-performance liquid chromatography (IP-RP-HPLC). Both the length and the diameter of the connecting capillaries (150 x 0.020 mm I.D.) as well as the detection volume (3 nl) had to be kept to a minimum in order to maintain the high efficiency of this chromatographic separation system with peak widths at half height in the range of a few seconds. Three different types of frits, namely sintered silica particles, sintered octadecylsilica particles, and monolithic poly(styrene-divinylbenzene) (PS-DVB) frits were evaluated with respect to their influence on chromatographic performance. Best performance for the separation of oligonucleotides and long DNA fragments was observed with the PS-DVB frits, whereas the short DNA fragments were optimally resolved in columns terminated by octadecylsilica frits. The maximum loading capacity of 60 x 0.20 mm I.D. columns ranged from 20 fmol (7.7 ng) for a 587 base pair DNA fragment to 500 fmol (2.4 ng) for a 16-mer oligonucleotide. Lower mass- and concentration detection limits in the low femtomol and low nanomol per liter range, respectively, make capillary IP-RP-HPLC with UV absorbance detection highly attractive for the separation and characterization of minute amounts of synthetic oligonucleotides, DNA restriction fragments, and short tandem repeat sequences amplified by polymerase chain reaction.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11043584&dopt=Abstract

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