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J Biol Chem. 2001 Jan 26;276(4):2354-60. Epub 2000 Oct 16.
2'-O,4'-C-methylene bridged nucleic acid modification promotes pyrimidine motif triplex DNA formation at physiological pH: thermodynamic and kinetic studies.

Torigoe H, Hari Y, Sekiguchi M, Obika S, Imanishi T.

Tsukuba Life Science Center, Institute of Physical and Chemical Research (RIKEN), Ibaraki 305-0074, Japan. torigotc.riken.go.jp

Extreme instability of pyrimidine motif triplex DNA at physiological pH severely limits its use in an artificial control of gene expression in vivo. Stabilization of the pyrimidine motif triplex at physiological pH is, therefore, crucial in improving its therapeutic potential. To this end, we have investigated the thermodynamic and kinetic effects of our previously reported chemical modification, 2'-O,4'-C-methylene bridged nucleic acid (2',4'-BNA) modification of triplex-forming oligonucleotide (TFO), on pyrimidine motif triplex formation at physiological pH. The thermodynamic analyses indicated that the 2',4'-BNA modification of TFO increased the binding constant of the pyrimidine motif triplex formation at neutral pH by approximately 20 times. The number and position of the 2',4'-BNA modification introduced into the TFO did not significantly affect the magnitude of the increase in the binding constant. The consideration of the observed thermodynamic parameters suggested that the increased rigidity itself of the 2',4'-BNA-modified TFO in the free state relative to the unmodified TFO may enable the significant increase in the binding constant at neutral pH. Kinetic data demonstrated that the observed increase in the binding constant at neutral pH by the 2',4'-BNA modification of TFO resulted from the considerable decrease in the dissociation rate constant. Our results certainly support the idea that the 2',4'-BNA modification of TFO could be a key chemical modification and may eventually lead to progress in therapeutic applications of the antigene strategy in vivo.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11035027&dopt=Abstract

Am J Obstet Gynecol. 2000 Oct;183(4):948-55.
Cervical shedding of herpes simplex virus and cytomegalovirus throughout the menstrual cycle in women infected with human immunodeficiency virus type 1.

Mostad SB, Kreiss JK, Ryncarz A, Chohan B, Mandaliya K, Ndinya-Achola J, Bwayo JJ, Corey L.

Departments of Epidemiology, Medicine, and Laboratory Medicine, University of Washington, Seattle, WA 98195, USA.

OBJECTIVE: Our purpose was to evaluate the frequency and patterns of the shedding of herpes simplex virus and cytomegalovirus in the female genital tract throughout the menstrual cycle. STUDY DESIGN: Seventeen women, all seropositive for herpes simplex virus types 1 and 2, cytomegalovirus, and human immunodeficiency virus type 1, underwent daily evaluation of cervical viral shedding for the duration of 1 menstrual cycle (21-31 visits per woman). Serum estradiol and progesterone levels were monitored 3 times weekly. RESULTS: Overall, herpes simplex virus deoxyribonucleic acid was detected in 43 (10%) of 450 cervical swabs, and cytomegalovirus deoxyribonucleic acid was detected in 232 (52%) of 450 cervical swabs. For individual women there was considerable variability in the percentage of days on which virus was detected, ranging from 0% to 33% for herpes simplex virus and from 20% to 97% for cytomegalovirus. Shedding of herpes simplex virus did not vary significantly with menstrual cycle; however, shedding of cytomegalovirus was significantly more frequent in the luteal phase (odds ratio, 1.9; 95% confidence interval, 1.1-3.4). A CD4(+) lymphocyte count <200/microL was associated with increased frequency of the detection of herpes simplex virus (odds ratio, 5.7; 95% confidence interval, 1.1-29.4). CONCLUSIONS: Asymptomatic cervical shedding of both herpes simplex virus and cytomegalovirus occurs very frequently in women infected with human immunodeficiency virus type 1. The risk of transmitting these viruses to sexual partners and neonates may be higher than previously recognized.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11035345&dopt=Abstract

Arch Pathol Lab Med. 2000 Oct;124(10):1510-3.
Natural killer-cell lymphoma involving the gynecologic tract.

Mhawech P, Medeiros LJ, Bueso-Ramos C, Coffey DM, Gei AF, Shahab I.

Departments of Pathology, University of Texas, M.D Anderson Medical Center, Houston, TX 77030, USA.

Non-Hodgkin lymphomas (NHL) can involve the gynecologic tract, most often as a manifestation of systemic involvement, and most cases reported have been of B-cell lineage. We describe 2 women with natural killer (NK)-cell lymphoma involving the gynecologic tract that initially presented with vaginal bleeding. In case 1, the patient had a stage IE nasal-type NK-cell lymphoma involving the cervix. The tumor was composed of medium-sized, irregular lymphoid cells with angioinvasion and necrosis. In case 2, the patient had a stage IV blastoid NK-cell lymphoma/leukemia infiltrating all organs in a hysterectomy and bilateral salpingo-oophorectomy specimen. Subsequent biopsy specimens revealed that the bone marrow and lymph nodes were also involved. The neoplasm was composed of small to medium lymphoid cells with fine nuclear chromatin. Case 1 was assessed immunohistochemically and the neoplastic cells were positive for CD3, CD56, and TIA-1. Case 2 was analyzed using both immunohistochemical and flow cytometry methods. The neoplastic cells were positive for cytoplasmic CD3, CD4, CD7, CD43, CD45, and CD56 and were negative for surface CD3. Both cases were negative for Epstein-Barr virus (EBV) ribonucleic acid (RNA) and molecular studies showed no evidence of T-cell receptor gamma chain gene rearrangements. The immunophenotype and absence of T-cell receptor gene rearrangements support NK-cell origin. We report these cases to illustrate that NK-cell lymphomas can involve, and rarely arise in, the gynecologic tract.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11035585&dopt=Abstract

Exp Cell Res. 2000 Nov 1;260(2):248-56. ["Cited in Books","window.top.location='/entrez/query.fcgi?tool=pmcited&cmd=search&db=books&term=11035919[pmid]'","",""],
Segregation of RNA and separate packaging of DNA and RNA in apoptotic bodies during apoptosis.

Halicka HD, Bedner E, Darzynkiewicz Z.

Brander Cancer Research Institute, Hawthorne, New York, 10532, USA.

Apoptosis is characterized by a complex and remarkably ordered choreography of events consisting of the preparatory and execution steps that all culminate in disposal of the cell remnants. The disposal occurs in a manner that is the least destructive to the tissue: the remains of nuclear chromatin and cytoplasm are packaged in apoptotic bodies which are then phagocytized by neighboring live cells without invoking inflammatory or autoimmune response. In the present study we describe that in the course of apoptosis cellular RNA becomes sequestered and packaged into granules and then into apoptotic bodies, separately from DNA. This separation, which appears to be initiated by the nucleolar segregation, was observed in HL-60 cells that were undergoing spontaneous apoptosis in cultures or were treated with the DNA-damaging drug, DNA topoisomerase I inhibitor camptothecin (CPT), or with the cell death ligand, tumor necrosis factor-alpha. RNA separation was also observed in apoptotic MCF-7 cells following treatment with CPT. RNA and DNA in apoptotic cells were identified histochemically, by their differential stainability with pyronin Y and Hoechst 33342 fluorochromes, respectively, and immunocytochemically, by labeling the RNA with BrU for various periods of time and detection of the incorporated precursor with fluoresceinated anti-BrU mAb; DNA was counterstained with 7-aminoactinomycin D. Over 90% of apoptotic bodies that contained RNA had no detectable DNA and vice versa, the apoptotic bodies containing DNA had no detectable RNA. Packaging RNA and DNA into separate apoptotic bodies suggests that the phagosomes of the cells that ingest these particles are specialized: some of them are responsible for DNA degradation, others for degradation of RNA. Such specialization may facilitate heterophagic degradation of nucleic acids during apoptosis. 2000 Academic Press.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11035919&dopt=Abstract

Gastrointest Endosc Clin N Am. 2000 Oct;10(4):573-93.
Molecular and immunologic pathology for the endoscopist. Special techniques.

Cortina G.

Department of Pathology and Laboratory Medicine, University of California Los Angeles, USA.

Special techniques for the endoscopic biopsy include both immunologic (IHC) and molecular approaches (ISH, PCR). Both approaches have been adapted to foster additional diagnostic power with little or no additional work by the endoscopist. The techniques take advantage of tissue-specific antigens, unique markers in infectious diseases, and unique nucleic acid sequences present in some malignancies. From these advantages, benign conditions may be more easily distinguished from malignant ones, the causes of some infectious diseases can be confirmed, and clinically relevant classification of malignancies can be made. Some special techniques have added difficulties, such as fastidious requirements for some IHC or Southern blotting. At UCLA, approaches with the least technical challenges are those in practice for endoscopic biopsy. Lastly, the information gleaned from special techniques requires endoscopic and histopathologic context for accurate interpretation.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11036534&dopt=Abstract

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