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At least 199 records · Page 11Linked to original sources
PubMed2000

Effects of body-mind training and relaxation stretching on persons with chronic toxic encephalopathy.

The purpose of this project was to investigate the psychological and physical effects of training of body awareness and slow stretching on persons with chronic toxic encephalopathy (CTE). In the present study, a method of self-regulation, a body-mind training, is presented. The body-mind training used was a guided relaxation technique combined with meditative stretching. The techniques are introduced and the psychological and physiological effects of the training is presented. Eight subjects with CTE, 48.5 years, were trained for 8 weeks. Outcome measures were percentage alpha brain waves (alpha%), electromyography (EMG) on the frontalis muscle, state-trait anxiety (STAI), creativity (RAT), and mood measured as anxiousness, humour and mental fatigue. The mean alpha% increased 52% during the training period (P < 0.01), and the EMG decreased 31% (P < 0.001. State anxiety decreased 22% during the training period (P < 0.01), but no changes were observed in trait anxiety and in the creativity score. The level of anxiousness and fatigue before a training session decreased during the training period. In conclusion, the body-mind training resulted in an improved ability for physical and mental relaxation as indicated from the lower EMG, the higher alpha% and the decrease in state anxiety.

Alpha Rhythm↗
PubMed1999

Characterization of unique DNA-binding and transcriptional-activation functions in the carboxyl-terminal extension of the zinc finger region in the human vitamin D receptor.

The vitamin D receptor (VDR) binds 1,25-dihydroxyvitamin D(3) and mediates its actions on gene transcription by heterodimerizing with retinoid X receptors (RXRs) on direct repeat (DR+3) vitamin D responsive elements (VDREs) located in target genes. The VDRE binding function of VDR has been primarily ascribed to the zinc finger region (residues 24-87). To define the minimal VDRE binding domain for human VDR (hVDR), a series of C-terminally truncated hVDR mutants (Delta134, Delta113, Delta102, Delta90, Delta84, Delta80, and Delta60) was generated and expressed in bacteria. Only the Delta134 and Delta113 mutants bound the VDRE (predominantly as monomers), suggesting that, in addition to the conserved zinc finger region of hVDR, as many as 25 amino acids in a C-terminal extension (CTE) participate in DNA binding. Site-directed mutagenesis of conserved charged residues in full-length hVDR was then performed to dissect the functional significance of the CTE (residues 88-112) in the context of the complete hVDR-RXR-VDRE interaction. Functional assays revealed that E98K/E99K, R102A/K103A/R104A, and K109A/R110A/K111A mutant hVDRs possessed dramatically reduced DNA binding and transcriptional activities, whereas distinct point mutants, such as K103A, bound to DNA normally but lacked transcriptional activity. Therefore, the boundary for the minimal DNA-binding domain in hVDR extends C-terminal of the zinc fingers to Lys-111, with clusters of highly conserved charged amino acids playing a crucial role in binding to the DR+3 element. Further, individual residues in this region (e.g., Lys-103) may lie on the opposing face of a DNA-binding alpha-helix, where they could contact transcriptional coactivators or basal transcription factors.

Amino Acid Sequence↗
PubMed2004

Proximal C-terminal domain of sulphonylurea receptor 2A interacts with pore-forming Kir6 subunits in KATP channels.

Functional KATP (ATP-sensitive potassium) channels are hetero-octamers of four Kir6 (inwardly rectifying potassium) channel subunits and four SUR (sulphonylurea receptor) subunits. Possible interactions between the C-terminal domain of SUR2A and Kir6.2 were investigated by co-immunoprecipitation of rat SUR2A C-terminal fragments with full-length Kir6.2 and by analysis of cloned KATP channel function and distribution in HEK-293 cells (human embryonic kidney 293 cells) in the presence of competing rSUR2A fragments. Three maltose-binding protein-SUR2A fusions, rSUR2A-CTA (rSUR2A residues 1254-1545), rSUR2A-CTB (residues 1254-1403) and rSUR2A-CTC (residues 1294-1403), were co-immunoprecipitated with full-length Kir6.2 using a polyclonal anti-Kir6.2 antiserum. A fourth C-terminal domain fragment, rSUR2A-CTD (residues 1358-1545) did not co-immunoprecipitate with Kir6.2 under the same conditions, indicating a direct interaction between Kir6.2 and a 65-amino-acid section of the cytoplasmic C-terminal region of rSUR2A between residues 1294 and 1358. ATP- and glibenclamide-sensitive K+ currents were decreased in HEK-293 cells expressing full-length Kir6 and SUR2 subunits that were transiently transfected with fragments rSUR2A-CTA, rSUR2A-CTC and rSUR2A-CTE (residues 1294-1359) compared with fragment rSUR2A-CTD or mock-transfected cells, suggesting either channel inhibition or a reduction in the number of functional KATP channels at the cell surface. Anti-KATP channel subunit-associated fluorescence in the cell membrane was substantially lower and intracellular fluorescence increased in rSUR2A-CTE expressing cells; thus, SUR2A fragments containing residues 1294-1358 reduce current by decreasing the number of channel subunits in the cell membrane. These results identify a site in the C-terminal domain of rSUR2A, between residues 1294 and 1358, whose direct interaction with full-length Kir6.2 is crucial for the assembly of functional KATP channels.

ATP-Binding Cassette Transporters↗
PubMed2001

RNA-protein hybrid ribozymes that efficiently cleave any mRNA independently of the structure of the target RNA.

Ribozyme activity in vivo depends on achieving high-level expression, intracellular stability, target colocalization, and cleavage site access. At present, target site selection is problematic because of unforeseeable secondary and tertiary RNA structures that prevent cleavage. To overcome this design obstacle, we wished to engineer a ribozyme that could access any chosen site. To create this ribozyme, the constitutive transport element (CTE), an RNA motif that has the ability to interact with intracellular RNA helicases, was attached to our ribozymes so that the helicase-bound, hybrid ribozymes would be produced in cells. This modification significantly enhanced ribozyme activity in vivo, permitting cleavage of sites previously found to be inaccessible. To confer cleavage enhancement, the CTE must retain helicase-binding activity. Binding experiments demonstrated the likely involvement of RNA helicase(s). We found that attachment of the RNA motif to our tRNA ribozymes leads to cleavage in vivo at the chosen target site regardless of the local RNA secondary or tertiary structure.

Base Sequence↗
PubMed1997

Mutations in the nuclear export signal of human ran-binding protein RanBP1 block the Rev-mediated posttranscriptional regulation of human immunodeficiency virus type 1.

We identified a region in the human Ran GTPase-binding protein RanBP1 that shares similarities to the nuclear export signal of the inhibitor of the cAMP-dependent protein kinase. Mutational analysis confirmed that this region is responsible for the cytoplasmic accumulation of RanBP1 and can functionally replace the nuclear export signal of Rev of human immunodeficiency virus type 1. We showed that RanBP1 interferes with Rev-mediated expression of human immunodeficiency virus type 1, whereas the RanBP1 with inactivated nuclear export signal abrogates Rev function. Expression of a Rev-independent molecular clone, which is regulated via the constitutive transport element (CTE) of the simian retrovirus type 1, is not affected. These findings indicate that Rev and RanBP1 compete for the same nuclear export pathway, whereas Rev- and the CTE-mediated pathways are distinct. The inhibition of Rev function is independent of the ability of RanBP1 to associate with Ran and therefore, it is not likely a result of interference with Ran function. These data suggest that RanBP1 interacts with Rev at the putative nuclear receptor and, hence, shares a step in posttranscriptional pathway with Rev.

Amino Acid Sequence↗
PubMed2000

Modified human immunodeficiency virus-based lentiviral vectors display decreased sensitivity to trans-dominant Rev.

As a first step toward the development of HIV-based conditionally replicating defective interfering particles expressing trans-dominant Rev (TdRev), we studied whether mutation of the splicing signals and replacement of the RRE by the SRV-1 CTE would render these vectors less sensitive to TdRev. Vectors with mutations in the splicing signals (SD-/RRE+) yielded high titers (5 X 10(6) CFU/ml) and showed higher levels of cytoplasmic unspliced mRNA than the corresponding SD+/RRE+ vectors either in the absence of Rev, in the presence of TdRev, or in the presence of both TdRev and Rev. Proviral copies of SD-/RRE+ vectors were rescued more efficiently than SD+/RRE+ vectors when TdRev was expressed. Vectors with the SRV-1 CTE (SD+/CTE+ and SD-/CTE+) expressed high levels of cytoplasmic unspliced mRNA in the absence of Rev expression. Titers obtained with the SD-/CTE+ vectors (10(6) CFU/ml) were higher than the titers obtained with SD+/CTE+ vectors. We also tested the effect of other structural modifications such as the orientation of the expression cassette and the presence of the central polypurine tract (cPPT/CTS). We show that an expression cassette cloned in the reverse orientation with respect to the LTRs or elimination of the cPPT/CTS element severely affected vector titers. We also demonstrated that these vectors can be efficiently mobilized from their proviral state by HIV trans-complementing functions, and transduced into secondary target cells without suffering any genomic rearrangement.

Base Sequence↗
PubMed2002

Enhancement of antibody responses to an HIV-2 DNA envelope vaccine using an expression vector containing a constitutive transport element.

Because immune responses to DNA vaccines in humans remains suboptimal, strategies need to be devised to facilitate expression of the vaccine in vivo. One method to improve response to a DNA vaccine is to construct plasmid vectors with leader sequences and post-transcriptional elements that facilitate export of transcribed RNA. In this study, we sought to determine if a mammalian expression vector (pND-14) containing a tissue plasminogen activator (TPA) leader sequence and a constitutive transport element (CTE) from simian retrovirus was superior to other mammalian expression vectors containing a post-transcriptional regulatory element (PRE) from hepatitis B virus (pCMV-link) or a minimal mammalian expression vector (pVAX1). Toward this objective, we evaluated protein expression of the HIV-2 envelope gene (gp140) in vitro and immune responses in immunized mice. We found that pVAX1 produced three- to fourfold lower levels of gp140 in vitro (5 ng/ml) in contrast to the pCMV-link and pND-14 vectors. When we immunized groups of mice intradermally with two of the HIV-2 gp140 DNA vaccine constructs, we found that pND-14 induced higher levels of envelope-specific systemic and mucosal antibodies than pCMV-link. We conclude that expression vectors for DNA vaccines should contain TPA and CTE sequences to facilitate immune responses.

AIDS Vaccines↗
PubMed2000

The structure of the mRNA export factor TAP reveals a cis arrangement of a non-canonical RNP domain and an LRR domain.

Human TAP is implicated in mRNA nuclear export and is used by simian type D retroviruses to export their unspliced genomic RNA to the cytoplasm of the host cell. We have determined the crystal structure of the minimal TAP fragment that binds the constitutive transport element (CTE) of retroviral RNAs. Unexpectedly, we find the fragment consists of a ribonucleoprotein (RNP) domain, which is not identifiable by its sequence, and a leucine-rich repeat (LRR) domain. The non-canonical RNP domain functions as the general RNA-binding portion of the fragment. The LRR domain is required in cis to the RNP domain for CTE RNA binding. The structural and biochemical properties of the domains point to a remarkable similarity with the U2B"(RNP)-U2A'(LRR) spliceosomal heterodimer. Our in vitro and in vivo functional studies using structure-based mutants suggest that a phylogenetically conserved surface of the LRR domain of TAP may have different roles in the export of viral and cellular RNAs.

Amino Acid Sequence↗
PubMed2006

The GapA/B gene duplication marks the origin of Streptophyta (charophytes and land plants).

Independent evidence from morphological, ultrastructural, biochemical, and molecular data have shown that land plants originated from charophycean green algae. However, the branching order within charophytes is still unresolved, and contradictory phylogenies about, for example,the position of the unicellular green alga Mesostigma viride are difficult to reconcile. A comparison of nuclear-encoded Calvin cycle glyceraldehyde-3-phosphate dehydrogenases (GAPDH) indicates that a crucial duplication of the GapA gene occurred early in land plant evolution. The duplicate called GapB acquired a characteristic carboxy-terminal extension (CTE) from the general regulator of the Calvin cycle CP12. This CTE is responsible for thioredoxin-dependent light/dark regulation. In this work, we established GapA, GapB, and CP12 sequences from bryophytes, all orders of charophyte as well as chlorophyte green algae, and the glaucophyte Cyanophora paradoxa. Comprehensive phylogenetic analyses of all available plastid GAPDH sequences suggest that glaucophytes and green plants are sister lineages and support a positioning of Mesostigma basal to all charophycean algae. The exclusive presence of GapB in terrestrial plants, charophytes, and Mesostigma dates the GapA/B gene duplication to the common ancestor of Streptophyta. The conspicuously high degree of GapB sequence conservation suggests an important metabolic role of the newly gained regulatory function. Because the GapB-mediated protein aggregation most likely ensures the complete blockage of the Calvin cycle at night, we propose that this mechanism is also crucial for efficient starch mobilization. This innovation may be one prerequisite for the development of storage tissues in land plants.

Amino Acid Sequence↗
PubMed1991

Continuous versus conventional tissue expansion: experimental verification of a new technique.

Historically, tissue expansion is a prolonged process, typically requiring at least 6 weeks to complete. Recently, interest has increased in shortening this time period. In the current study, a continuous infusion device maintaining constant expander pressure less than capillary filling pressure was used in a canine model in seven dogs to minimize the time period needed to achieve significant expansion. There were no complications, except one device malfunction, corrected by changes in design. The process was shown to be a safe and effective means of producing amounts of expansion similar to traditional methods in approximately 72 hours, with expansion of 28 percent (n = 6) for continuous tissue expansion (CTE) versus 34 percent (n = 6) for a 2-week rapid expansion protocol. This expansion was derived from either stretch of preexisting tissue (46 percent for CTE, 35 percent for 2-week expansion) or recruitment of adjacent tissue. The clinical application of continuous tissue expansion could permit the advantages of tissue expansion to be obtained in many more situations than are currently available to traditional tissue expansion techniques.

Animals↗
PubMed1981

Evaluation of antithymocyte globulin for human bone marrow transplantation. III. Effect of antithymocyte globulin treatment on t cell precursors.

The effect of antithymocyte globulin (ATG) treatment on T cell precursors was investigated in an in vitro model of E rosette induction by thymic factors. Approximately 8% of bone marrow mononuclear cells can be induced to form E rosettes following incubation with thymosin or with conditioned medium from cultured thymic epithelium (CM-CTE). Treatment of bone marrow mononuclear cells with ATG alone markedly reduced but did not eliminate precursors of E rosette-forming cells (E-RFCs), whereas treatment with ATG and complement completely inhibited these bone marrow E-RFC precursors. E-RFCs already present in peripheral blood and bone marrow were completely inhibited by treatment with ATG alone. E-RFCs induced by treatment of bone marrow cells with thymosin or CM-CTE were also completely inhibited by ATG alone. These data indicate that bone marrow precursors of E-RFCs are less sensitive to ATG inhibition than are peripheral blood and bone marrow E-RFCs. When bone marrow precursor T cells are induced to become E-RFCs by the treatment with thymic factors, they acquire comparable sensitivity to ATG. A similar disparity in sensitivity to ATG treatment was observed when ATGs absorbed with human fetal liver cells or granulocytes were studied. These data suggest that ATG treatment of bone marrow cells may have different effects on mature and precursor T cells. It is important to consider these factors in attempts to use ATG treatment in clinical bone marrow transplantation trials.

Antilymphocyte Serum↗
PubMed2006

Correlation transfer and diffusion of ultrasound-modulated multiply scattered light.

We develop a temporal correlation transfer equation (CTE) and a temporal correlation diffusion equation (CDE) for ultrasound-modulated multiply scattered light. These equations can be applied to an optically scattering medium with embedded optically scattering and absorbing objects to calculate the power spectrum of light modulated by a nonuniform ultrasound field. We present an analytical solution based on the CDE and Monte Carlo simulation results for light modulated by a cylinder of ultrasound in an optically scattering slab. We further validate with experimental measurements the numerical calculations for an actual ultrasound field. The CTE and CDE are valid for moderate ultrasound pressures and on a length scale comparable with the optical transport mean-free path. These equations should be applicable to a wide spectrum of conditions for ultrasound-modulated optical tomography of soft biological tissues.

Animals↗
PubMed2005

Regulation of photosynthetic GAPDH dissected by mutants.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of higher plants catalyzes an NADPH-consuming reaction, which is part of the Calvin cycle. This reaction is regulated by light via thioredoxins and metabolites, while a minor NADH-dependent activity is constant and constitutive. The major native isozyme is formed by A- and B-subunits in stoichiometric ratio (A2B2, A8B8), but tetramers of recombinant B-subunits (GapB) display similar regulatory features to A2B2-GAPDH. The C-terminal extension (CTE) of B-subunits is essential for thioredoxin-mediated regulation and NAD-induced aggregation to partially inactive oligomers (A8B8, B8). Deletion mutant B(minCTE) is redox insensitive and invariably tetrameric, and chimeric mutant A(plusCTE) acquired redox sensitivity and capacity to aggregate to very large oligomers in presence of NAD. Redox regulation principally affects the turnover number, without significantly changing the affinity for either 1,3-bisphosphoglycerate or NADPH. Mutant R77A of GapB, B(R77A), is down-regulated and mimics the behavior of oxidized GapB under any redox condition, whereas mutant B(E362Q) is constantly up-regulated, resembling reduced GapB. Despite their redox insensitivity, both B(R77A) and B(E362Q) mutants are notably prone to aggregate in presence of NAD. Based on structural data and current functional analysis, a model of GAPDH redox regulation is presented. Formation of a disulfide in the CTE induces a conformational change of the GAPDH with repositioning of the terminal amino acid Glu-362 in the proximity of Arg-77. The latter residue is thus distracted from binding the 2'-phosphate of NADP, with the final effect that the enzyme relaxes to a conformation leading to a slower NADPH-dependent catalytic activity.

Amino Acid Sequence↗
PubMed2005

Moxifloxacin-based strategies for first-line treatment of Helicobacter pylori infection.

BACKGROUND: Standard anti-Helicobacter pylori therapy may not achieve a satisfactory eradication rate. Fluoroquinolones, such as moxifloxacin, are safe and promising agents for H. pylori eradication. AIM: To compare the efficacy of two 1-week moxifloxacin-based H. pylori eradication regimens with two standard treatments. METHODS: Three hundred and twenty H. pylori-positive subjects were randomized into four groups to receive: moxifloxacin, amoxicillin, esomeprazole (Group MAE); moxifloxacin, tinidazole and esomeprazole (Group MTE); standard triple therapies with clarithromycin, amoxicillin and esomeprazole (Group CAE) or tinidazole (Group CTE) for 7 days. H. pylori status was re-assessed 6 weeks after the end of therapy by 13C urea breath test. RESULTS: Three hundred and twenty patients completed the efficacy analysis per protocol; H. pylori eradication rate in group MTE was 90% (72 of 80) and 92% (72 of 78), in group MAE was 88% (70 of 80) and 89%, (70 of 79) in Group CAE was 73% (58 of 80) and 78% (58 of 74), and in Group CTE was 75% (60 of 80) and 79% (60 of 76), respectively, in intention-to-treat and in per protocol analyses. Eradication rates of moxifloxacin-based triple therapies were significantly higher than that observed using standard triple schemes. The incidence of side effects was significantly lower in moxifloxacin groups than in control groups. CONCLUSIONS: Seven-day moxifloxacin-based triple therapies provide optimal eradication rates with a good compliance when compared with the standard triple therapy schemes.

Adult↗
PubMed1994

Optical recordings of ventricular excitability of frog heart by an extracellular stimulating point electrode.

To enhance understanding of the excitability of cardiac muscle during rest, an optical technique using the fluorescent voltage sensitive dye di-4-ANEPPS was used. Unlike conventional electrical recordings, optical recordings are free from electrical artifacts and, therefore, allow the observation of the transmembrane potential not only following the stimulation pulse, but also during the pulse itself. Transmembrane potentials (Vm) were recorded optically from frog ventricular epicardium in calcium containing Ringer's solution directly under an extracellular stimulating point electrode. Anodal and cathodal S1 stimuli were applied at rest. As observed by previous investigators, the post-pulse excitatory responses for cathodal pulses, compared with anodal pulses were greater. Changes in transmembrane potential (delta Vm) during the pulse were as expected for a passive cable only for low intensity pulses (< 4 x the cathodal threshold of excitation in diastole, CTE). However, at the higher intensities necessary to produce an excitatory response (> 6-8 x CTE), an "irregular" response in Vm was observed--a reversal of the hyperpolarization during an anodal stimulus pulse and a reversal of the depolarization during a cathodal stimulus pulse. To elucidate further the biophysical basis for this behavior, delta Vm was mapped around the stimulating electrode. During stimulation, regions could be observed having a response with opposite polarity to that under the electrode (i.e., depolarization for an anodal pulse and hyperpolarization for a cathodal pulse). Removal of the bath solution or the addition of channel blockers did not eliminate the occurrence of these regions. These regions appear to be the basis for the irregular behavior of delta Vm directly under the electrode as well as for anodal excitation.

Animals↗
PubMed1992

High frequency of telomeric associations and chromatid exchanges and breaks in human ovarian carcinoma.

Cytogenetic analysis of an unselected series of twenty human serous adenocarcinomas and undifferentiated carcinomas of the ovary revealed the presence of telomeric associations (tas) and unstable chromosomal aberrations, including chromatid-type exchanges (cte) and breaks (ctb) in high proportion of tumors studied. Tas and cte & ctb were present in 75% and 55% of tumors, respectively. Involvement of different chromosome telomeres in tas seemed to be random. This is the first report describing telomeric associations in ovarian cancer. Our findings suggest that the frequency of extensive spontaneous chromosome breakage in ovarian carcinoma may be higher than that reported in the literature.

Adult↗
PubMed2005

Hairpin ribozymes in combination with siRNAs against highly conserved hepatitis C virus sequence inhibit RNA replication and protein translation from hepatitis C virus subgenomic replicons.

Chronic hepatitis C virus (HCV) infection is a clinically important liver disease with limited therapeutic options in a significant proportion of patients. Therefore, novel efficient therapeutic agents are needed. Because the 5'- and 3'-untranslated regions (UTRs) of HCV are highly conserved and functionally important for HCV replication, they are attractive targets for RNA-cleaving ribozymes or small interfering RNAs (siRNAs). In this study hairpin ribozymes (Rz) targeting HCV 5'- and 3'-UTR sequences were expressed from a retroviral vector transcript under control of two different RNA polIII promoters (tRNA(Val), U6). Ribozymes were evaluated in monocistronic, subgenomic I389/hyg-ubi/NS3-3'/5.1 HCV replicon cells as single agents or in combination with siRNAs against HCV 5'- or 3'-UTR recently demonstrated to inhibit HCV replicons. Additionally, ribozyme constructs were generated with the 3'-terminus of the ribozyme flanked by constitutive transport element (CTE) sequences, an RNA motif that has previously been shown to enhance cleavage activity of hammerhead ribozymes. In our study, tRNA(Val) as well as U6 promoter-driven Rzs markedly reduced HCV replicon RNA expression and HCV internal ribosome entry site (IRES)-mediated HCV NS5B protein translation from monocistronic subgenomic replicons. However, attachment of CTE sequences to the 3'-terminus did not significantly enhance activity of Rzs tested in this study. Interestingly, we detected additive HCV inhibitory effects for combinations of tRNA(Val)-driven Rzs and U6-derived siRNAs both directed against highly conserved 5'- and 3'-UTR sequence, suggesting that a dual strategy of ribozymes and siRNAs might become a powerful molecular tool to specifically silence HCV RNA replication.

Blotting, Western↗
PubMed1997

The effect of viral regulatory protein expression on gene delivery by human immunodeficiency virus type 1 vectors produced in stable packaging cell lines.

We describe the generation of stable human immunodeficiency virus type 1 (HIV-1)-packaging lines that constitutively express high levels of HIV-1 structural proteins in either a Rev-dependent or a Rev-independent fashion. These cell lines were used to assess gene transfer by using an HIV-1 vector expressing the hygromycin B resistance gene and to study the effects of Rev, Tat, and Nef on the vector titer. The Rev-independent cell lines were created by using gag-pol and env expression vectors that contain the Mason-Pfizer monkey virus (MPMV) constitutive transport element (CTE). Vector titers approaching 10(4) CFU/ml were routinely obtained with these cell lines, as well as with the Rev-dependent cell lines, with HeLa-CD4 cells as targets. The presence of Nef and Tat in the producer cell each increased the vector titer 5- to 10-fold. Rev, on the other hand, was absolutely essential for gene transfer, unless the MPMV CTE was present in the vector. In that case, by using the Rev-independent cell lines for packaging, Rev could be completely eliminated from the system without a reduction in vector titer.

Base Sequence↗