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T Vo

Publications and source records attributed to T Vo.

15 recordsLinked to original sources

Identification and expression of a mouse ortholog of A2BP1.

Human ataxin-2 contains a polyglutamine repeat that is expanded in patients with spinocerebellar ataxia type 2 (SCA2). Ataxin-2 is highly conserved in evolution with orthologs in mouse, Caenorhabditis elegans, and Drosophila melanogaster. It interacts at its C-terminus with ataxin-2 binding protein 1, A2BP1. This study presents a highly conserved mouse ortholog of A2BP1, designated A2bp1. The amino acid sequence of the human and mouse protein is 97.6% identical. This remarkable degree of conservation supports the fact that these proteins have an important basic function in development and differentiation. Sequence analysis reveals the existence of RNA binding motifs. The A2bp1 transcript was found in various regions of the CNS including cerebellum, cerebral cortex, brain stem, and thalamus/hypothalamus. The A2bp1 protein was detected by immunocytochemistry in the CNS and connective tissue of the mouse embryo starting at stage E11, as well as in the heart at all stages. Mouse embryos showed varying expression of A2bp1 at all stages. Previous studies in other model systems had implicated the orthologs of ataxin-2 and A2BP1 in development. This study suggests a role for A2bp1 in embryogenesis as well as in the adult nervous system, possibly mediated by a function in RNA distribution or processing.

Animals↗

Should ultrasound criteria of the prostate be redefined to better evaluate when and where to biopsy.

During the past 10 years, it has been suggested, and accepted by some, that transrectal ultrasound (TRUS) of the prostate should be used to identify a hypoechoic lesion or, if needed, guide biopsy into nonspecific areas. Retrospectively, the authors attempted to evaluate the need to identify areas that were on pathologic analysis, prostate cancer, but were not hypoechoic, but would require random/systematic biopsy to exclude prostate cancer. Six-hundred fifteen consecutive men were referred to the authors because of a concern found on digital rectal examination or because of increase in prostate-specific antigen. All patients underwent TRUS-guided biopsy of the prostate using either the four-quadrant or sextant biopsy technique. Each area undergoing biopsy was characterized as: 1) normal-appearing; 2) hypoechoic; 3) mixed echogenic (containing both hypoechoic and hyperechoic elements); 4) subtly hyperechoic (containing no calculi); or 5) isoechoic (lesion was seen because of distortion of the normal architecture). A diagnosis of carcinoma was made in 197 patients (32%). Of these, 99 (50.2%) patients had a hypoechoic lesion as the primary site, corresponding to their highest Gleason grade. Twenty-five (12.7%) had mixed echogenicity, nine (4.6%) had hyperechoic foci, and 23 (11.7%) had isoechoic biopsy-proven foci of prostate cancer. Forty-one (20.8%) patients with adenocarcinoma had normal ultrasound findings. The median Gleason grade for cancer in visible mixed echogenic and hyperechoic areas were generally higher than that for cancer in hypoechoic sites. Hypoechoic cancer sites had a Gleason grade range of 2 to 10 (median 5); mixed echogenic foci had a Gleason range of 2 to 10 (median 6); hyperechogenic cancers had a Gleason range of 2 to 8 (median 6); isoechoic cancers had a Gleason range of 2 to 7 (median 5); normal foci had a Gleason range of 2 to 8 (median 5). Results of this study suggest that 50% of clinically significant prostate cancers are not purely hypoechoic, and 37% of all diagnosed cancers contain no hypoechoic elements.

Journal Article↗

Field trials of three concentrations of Laginex as biological larvicide compared to Vectobac-12AS as a biocontrol agent for Culex quinquefasciatus.

Laginex AS Biological Larvicide (Lagenidium giganteum) and Vectobac-12AS (Bacillus thuringiensis var. israelensis) were compared in plastic pools containing laboratory-reared Culex quinquefasciatus larvae located in a wooded area in Panama City, FL, in August and September 1997. The pools were dipped at 2-day intervals to assess larval control. Sentinel larvae in screen-sided cups were added to all treatment and control pools at 4-day intervals and evaluated for infection at 48 h. Laginex produced larval control up to 20 days as compared to the Vectobac-12AS compound, which required retreatment by the 10th day. The most effective control was attained by the Laginex 25 treatment, which reduced larvae in the pools by 100% for 22 days. Control in the Laginex 15 and Laginex 35 treatments dropped to 90% and 74% at day 13 and day 9, respectively. The numbers of infected larvae remained above 50% as larval control was assessed, but dropped and fluctuated when water temperatures were lowered at the onset of cold weather. Organic pollution did not prove to be a significant factor in the evaluations.

Animals↗

Prenatal diagnosis of 46,XY/46,XX mosaicism: a case report.

We report on the prenatal diagnosis of a fetus with 46,XY and 46,XX cell lines with a normal male phenotype. Cytogenetic and molecular studies ruled out the possibility of maternal cell contamination and showed that all the X chromosomes present in both fetal cell lines were derived from a single maternal X chromosome. This suggests 46,XY/46,XX mosaicism.

Alleles↗

Phosphorylation of TRAF2 inhibits binding to the CD40 cytoplasmic domain.

TRAF2 is a signal transducing adaptor molecule which binds to the CD40 cytoplasmic domain. We have found that it is phosphorylated, predominantly on serine residues, when transiently overexpressed in 293 cells. The phosphorylation appears to be related to the signaling events that are activated by TRAF2 under these circumstances, since two nonfunctional mutants were found to be phosphorylated significantly less than the wild-type protein. Furthermore, the phosphorylation status of TRAF2 had significant effects on the ability of the protein to bind to CD40, as evidenced by our observations that the CD40 cytoplasmic domain interacted preferentially with underphosphorylated TRAF2 and that phosphatase treatment significantly enhanced the binding of TRAF2 to CD40. We conclude from these studies that the phosphorylation of TRAF2 is likely to play an important role in regulating signaling by virtue of its ability to influence the CD40-TRAF2 interaction.

Amino Acid Substitution↗

Identification and cloning of a brain autoantigen in neuro-behavioral SLE.

In murine models of SLE, particular patterns of abnormalities of social interaction and memory collectively known as neurobehavioral dysfunction (NBD) correlate with the occurrence of brain reactive autoantibodies. Study of the immunopathogenic effects of these antibodies has been limited by the absence of isolated autoantibodies and antigens. In order to identify the molecular targets, we isolated autoantibodies highly specific for brain plasma membranes from MRL/lpr mice. After immunoscreening a brain expression library with these brain specific autoantibodies, we identified a single cDNA clone of unique sequence and relevant anatomic distribution. Transcript for this cDNA is wide spread among mammalian species but appears to be present only in the brain. Addition features, suggesting this cDNA is pertinent for further study include (1) the expressed protein, called lupus brain antigen 1, reacts with the screening immunoglobulins as well as immunoglobulins from other strains of murine neuro-SLE not used to screen the library, but not with immunoglobulins from normal mice, (2) the transcript distribution within the brain is similar to immunochemical localization of binding of the spontaneous autoantibodies and (3) the localization of transcript within the brain, in the hippocampus, hypothalamus an cingulate gyrus, corresponds to anticipated anatomical regions of clinical dysfunction. Further, the transcript is a large, potentially structural molecule of unique sequence. Antibodies to this molecule may mediate changes in behavior either by direct interactions with the cognate antigen or by indirect influences through neuro-endocrine axes.

Amino Acid Sequence↗

Evaluation of high-flux hemodiafiltration efficiency using an on-line urea monitor.

On-line urea monitoring of the effluent dialysate offers a real-time assessment of dialysis efficiency and metabolic/nutritional characteristics of hemodialysis patients. Quantitative parameters were evaluated by dialysate urea kinetic modeling (DUKM) with an on-line urea sensor in 23 patients treated by high-flux hemodiafiltration (HDF) (215 sessions of 210 to 240 minutes with a mean blood flow rate of 367 +/- 44 mL/min). Overall, the mean effective Kt/V was 1.52 +/- 0.29, the urea mass removed (22.8 +/- 5.5 g/session or 814 +/- 198 mmol/session), the solute removal index (SRI) 73% +/- 6.1%, and the mean normalized protein catabolic rate (nPCR), 1.15 +/- 0.31 g/kg/day. Blood urea kinetic modeling (BUKM), based on pre- and postsession urea concentrations, using equations from Daugirdas and Garred to calculate equilibrated Kt/V and nPCR, respectively, were in good agreement with DUKM, the differences observed appearing not clinically relevant. The variability of evaluated parameters was verified over consecutive sessions for a mean period of 3 weeks in the entire group. Mean variation in Kt/V was 8%; in urea mass removal, 18%; and in nPCR, 18%. When assessed over 1 week in a subgroup of 13 patients, Kt/V and PCR remained relatively stable, and urea mass removal per- and postsession declined from 23.5 +/- 8.0 g (840 +/- 285 mmol) to 18.7 +/- 6.3 g (667 +/- 225 mmol) from the first to the third session of the week, most likely as a consequence of interdialytic intervals. Predialysis urea concentrations followed the same trend. In the current study, DUKM with on-line urea sensor has confirmed that HDF is a highly efficient renal replacement modality; the variability observed in quantitative parameters supports a need for frequent adequacy monitoring. On-line urea monitoring of effluent dialysate is a simple device that provides the opportunity to tailor treatment to patient needs.

Blood Urea Nitrogen↗

Variable subcellular localization of a neuron-specific protein during NTera 2 differentiation into post-mitotic human neurons.

The current report describes the molecular characterization of the human (the D4S234 locus) and mouse (the m234) homologs of a gene that was isolated during our genomic analysis of the Huntington disease gene region. Sequence comparisons of full-length cDNA clones revealed that the mouse and human homologs encoded evolutionarily conserved 21-kDa proteins with greater than 90% amino acid sequence identity. Extensive sequence identity between the D4S234 gene and the rat p1A75 gene (a previously identified rat neuron-specific gene) showed that these genes are interspecies homologs. Furthermore, the D4S234 protein exhibited significant amino acid similarity to a 19-kDa mouse protein that localizes to the Golgi apparatus of embryonic neurons. However, nonconservative sequence differences suggested that these genes are independent members of a multigene family. Northern analyses revealed that rodent D4S234 expression occurred predominantly in the brain and included all brain regions. Neuron-specific expression was demonstrated using Northern analysis of cultured glial cells and quinolinic acid-treated rat brain samples. Minimal amounts of the rodent D4S234 mRNA were detected prenatally; however, elevated adult levels were detected within 1 month of birth. Sequence analyses of the human and mouse D4S234 proteins identified an evolutionarily conserved hydrophobic sequence and a consensus nuclear localization signal in both genes. Immunofluorescence microscopy, using an antipeptide antibody, established that the human D4S234 protein preferentially localized to the nucleus of mitotic cultured cells. Since the rat p1A75 protein was previously mapped to the neuronal cytoplasm by in situ hybridization, the subcellular localization of the D4S234 protein was subsequently examined during differentiation of the NTera 2 (NT2) cell line. Following differentiation into postmitotic NT2-N neurons, the D4S234 protein demonstrated cytoplasmic staining and reduced or undetectable nuclear staining in many cells. The variation in the intracellular localization of the D4S234 protein in mitotic and nonmitotic cells suggests that the subcellular localization of this protein is developmentally regulated and provides clues about the biochemical function of this protein.

Amino Acid Sequence↗

Role of Borrelia burgdorferi in the pathogenesis of morphea/scleroderma and lichen sclerosus et atrophicus: a PCR study of thirty-five cases.

Morphea (localized scleroderma), and lichen sclerosus et atrophicus (LSA) share common features with acrodermatitis chronica atrophicans (ACA), a known chronic form of borreliosis. These include similar histologic findings such as diffuse dermal fibrosis. These observations have led several investigators to consider the possibility of Borrelia burgdorferi (Bb) as a common etiologic factor among all of these diseases. The aim of this study is to investigate the role of Bb in the pathogenesis of morphea and LSA, by assaying for its presence in lesional skin biopsies from patients with these diseases. We utilized the nested polymerase chain reaction (PCR) technique to selectively amplify a longer segment of a Bb-specific somatic gene, on DNA from paraffin-embedded, formalin-fixed tissues. The results revealed no Bb-specific DNA sequence in 28 specimens of morphea/scleroderma and 7 of LSA with varying stages of disease. Furthermore, confirmatory Southern blot of the PCR product, resulted in similar findings. These data seriously question the role played by this spirochete in the pathogenesis of morphea and LSA, at least in the southeastern part of the USA.

Adult↗

Establishment of a human B-CLL xenograft model: utility as a preclinical therapeutic model.

Chronic lymphocytic leukemia (CLL), a proliferative disease of mature looking B lymphocytes, is the commonest leukemia in western countries. It remains incurable by available treatment modalities. We report on the establishment of a permanent, EBV-negative, B-CLL line (WSU-CLL) from the peripheral blood of a patient with CLL. The cells grow as suspension in liquid culture, express IgG lambda and other B cell markers and show lg heavy and light gene rearrangements. Karyotypic analysis shows 45,X,del(3)(p14;p24),t(4;12;12) (q31;q22;p13), t(5;12) (q31;p13), add(16)(q24)X2, t(18;21) (q12;p12). WSU-CLL forms colonies when grown on soft agar. A xenograft model was established by injecting the WSU-CLL cells subcutaneously (s.c.) in severe combined immune deficient (SCID) mice. When the s.c. tumor was transplanted in vivo to other SCID mice, the success rate was 100% with a doubling time of 7.3 days. The CLL-SCID xenograft model was used to test the efficacy of selected standard chemotherapy drugs and new therapeutic agents against WSU-CLL. The cell line and the xenograft described can be used as a model to facilitate the development of new therapeutic agents against CLL in man.

Aged↗

Targeting pyrimidine single strands by triplex formation: structural optimization of binding.

Recent reports describe a new strategy for the binding of single-stranded pyrimidine sequences by triple helix formation. In this approach, a double-length purine-rich oligonucleotide binds a target strand, folding back to form an antiparallel pur.pur.pyr triple helix. We now describe a series of studies in which sequence and structural variations are made in such purine-rich ligands, in an effort to optimize binding properties. Comparison is made between the use of two separate strands and the use of single two-domain ligands; the latter are found to bind more tightly and to aggregate less in media containing Na+ or K+. Placement of mismatched bases in the target shows that sequence selectivity of binding is as high as that for Watson-Crick duplex formation. Variation of the lengths and sequences of loops bridging the binding domains demonstrates that dinucleotide loops composed of pyrimidines give the highest stability. Oligoethylene glycol-derived loop replacements are shown to give good binding affinity as well. The binding of an RNA target is shown to occur with the same affinity as the binding of DNA. In general, it is found that circular variants bind more tightly than do either separate strands or singly-linked ligands and unlike linear oligomers, the circular compounds do not aggregate to a large extent even in buffers containing 100 mM K+. Such structurally optimized ligands are useful in expanding the number of possible naturally-occurring sequences which can be targeted by triplex formation.

Base Sequence↗

A Sau3A polymorphism in the 5' end of the IT15 gene that nonrandomly segregates with the Huntington disease trinucleotide expansion.

Genomic clones encompassing the Huntington disease (HD) mutation were used to isolate a probe that detects size changes in the restriction fragments that contain the HD trinucleotide repeat (TNR). This probe also detects a frequent Sau3A polymorphism (allele sizes 1.8-kb and 2.7kb), which maps approximately 950bp from the TNR. Examination of a number of HD families established that the frequency of the Sau3A alleles did not differ significantly between control and HD populations; however, the HD expansion was always present on a chromosome that contained the 1.8-kb Sau3A allele. This association between a specific allele and the HD TNR expansion was significant and could provide a clue to the chromosomal elements that produce the trinucleotide expansion on the Huntington disease chromosome.

Alleles↗

An estimate of the number of genes in the Huntington disease gene region and the identification of 13 transcripts in the 4p16.3 segment.

Physical mapping and genetic linkage studies have positioned the Huntington disease (HD) gene to a relatively large genomic region in the distal portion of the short arm of human chromosome 4 (4p16.3). To estimate the number of genes present in this region and to identify candidate disease genes, several clones that map to the 4p16.3 segment have been examined for clusters of CpG-rich restriction sites and transcribed sequences. Thirteen expressed sequences were identified and were shown by pulsed-field gel electrophoresis not to cluster into a small segment of the 4p16.3 band. The frequency of transcripts in these clones suggests that the putative HD gene region contains about 100 genes.

Blotting, Northern↗

In vitro evaluation of the Thermafil technique with and without gutta percha coating.

To determine the sealing ability of the Thermafil technique with and without gutta percha on the plastic carrier, 40 maxillary incisors were divided into four treatment groups: (A) Thermafil with gutta percha on the carrier; (B) Thermafil without gutta percha on the carrier; (C) lateral condensation with standardize gutta percha; and (D) a single standardized gutta percha cone. Average leakage in groups A-D were 0.64 mm, 1.32 mm, 0.53 mm, and 0.72 mm, respectively. The only significant difference was between the Thermafil without gutta percha and the other three groups (p < 0.05).

Analysis of Variance↗