Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Permits”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 559 records · Page 31Linked to original sources
PubMed1999

Direct proteasome inhibition by clasto-lactacystin beta-lactone permits the detection of ubiquitinated p21(waf1) in ML-1 cells.

The ubiquitin proteasome pathway regulates the expression of major cellular regulatory proteins. The ubiquitin proteasome system has been demonstrated to be involved in the expression of the cyclin kinase inhibitor, p21. Ubiquitinated p21 is degraded immediately by 26S proteasome, therefore, the detection of p21 is difficult. We report here an improvement for the detection of ubiquitinated p21 using a proteasome inhibitor, clasto-lactacystin beta-lactone. A p21-enriched cell lysate is obtained by pretreating the cells with deferoxamine to induce p21 mRNA expression followed by treatment with 1x10(-6) M beta-lactone. The concentration of p21 from the cell lysate was performed using an anti-p21 antibody crosslinked to protein G Sepharose. Ubiquitinated p21 was detected on Western blots of the concentrated sample using an anti-ubiquitin antibody. This detection system will be used for further analysis of the regulation of p21 ubiquitination.

Acetylcysteine↗
PubMed2003

A technique permitting selection of penicillin-susceptible pneumococcal strains following transformation of a penicillin-resistant strain with a penicillin-susceptibility gene.

We report a technique useful for transformation experiments involving bacteria naturally competent for DNA transformation. It allows the selection of antibiotic-susceptible transformants following the transformation of a resistant strain with an antibiotic susceptibility gene. We show the effectiveness of this technique through the selection of penicillin-susceptible (MIC, 0.03 microg/ml) transformants following the transformation of a penicillin-resistant (MIC, 16 microg/ml) pneumococcal strain with a penicillin-susceptibility gene.

Carrier Proteins↗
PubMed1986

Use of magnesium hydroxide and low magnesium dialysate does not permit reduction of aluminum hydroxide during continuous ambulatory peritoneal dialysis.

In an effort to reduce the ingestion of aluminum in phosphate-binding antacids, we treated seven patients on continuous ambulatory peritoneal dialysis (CAPD) with low magnesium dialysate and phosphate binders containing both aluminum and magnesium hydroxide. The total amount of phosphate binders prescribed was adjusted to maintain the serum phosphorus at normal levels. The dose of magnesium hydroxide was limited by intolerable gastrointestinal side effects in six of the seven patients. One patient also developed symptomatic hypermagnesemia. When magnesium hydroxide was prescribed in tolerable doses, the mean aluminum dose was not significantly decreased compared with the dose when taking aluminum hydroxide alone. We conclude that substitution of magnesium hydroxide for aluminum hydroxide as a phosphate binder fails to reduce the dose of aluminum in most patients on CAPD.

Adult↗
PubMed2001

Solvent interconnectedness permits measurement of proximal as well as distant phase transitions in polymer mixtures by fluorescence.

We monitored the fluorescence intensity and anisotropy of 1,6-diphenyl-1,3,5-hexatriene (DPH) incorporated in bovine serum albumin (BSA) and dimyristoylphosphatidylcholine (DMPC) vesicle membranes, which in turn were embedded in optically clear gelatin solutions, as a function of temperature. DPH in BSA gave unanticipated large changes in fluorescence intensity and anisotropy at the instant of gelatin gel melting. Both steady state anisotropy and fluorescence intensity reported the gel-sol transition point in gelatin unambiguously, which was independently confirmed as physical-pour point of the gel. In the case of DMPC vesicles, fluorescence intensity indicated the gelatin transition, while the anisotropy indicated DMPC phase transition. This fluorescence methodology uniquely offered a common probe for two distinct transitions in two distinct domains interconnected by the solvent, water.

Dimyristoylphosphatidylcholine↗
PubMed2001

Negative staining permits 4.0 A resolution with low-dose electron diffraction of catalase crystals.

Low-dose electron diffraction of thin single crystals of catalase that are negatively stained with the light-atom compound, dipotassium glucose-1,6-diphosphate, reveals Bragg reflections extending to 4.0A (= 0.40 nm). Under the same conditions, negative staining with the traditional heavy-metal salt, ammonium molybdate, also gives diffraction spots extending to 4.0 A. These results establish that negative staining of protein crystals preserves periodic structural information into the high-resolution range, unlike the widely accepted current belief that this methodology can give a resolution limited to only 20-25 A.

Catalase↗
PubMed2001

Primary culture of rat taste bud cells that retain molecular markers for taste buds and permit functional expression of foreign genes.

Taste buds are constituted of several kinds of cells which have distinct characteristics and play different roles. In this study, we have established an in vitro culture system by optimizing the method for isolating the cells and by selecting culture media and reagents effective for cell viability and adhesion. As a result, the taste bud cells were adhesive and viable for over 3 days when cultured onto Matrigel-coated dishes in medium based on keratinocyte growth medium. The cells retained molecular markers for both the cytoskeleton and intracellular signaling such as cytokeratin 8 and phospholipase Cbeta2. In addition, three intracellular signaling molecules, gustducin, phospholipase Cbeta2, and inositol 1,4,5-trisphosphate receptor type 3, are expressed in the same correlation as those in vivo, although the ratio of signaling molecule-positive cells vs. total cells was somewhat lower in the culture than in vivo. Next, we tried several methods to introduce foreign genes into the cells, and obtained a greater than 90% efficiency of introduction using an adenovirus vector. Finally, we show that an exogenously expressed myc-tagged alpha1A-adrenoceptor sorts into the plasma membrane, and transduces a ligand-dependent signal resulting in intracellular [Ca(2+)] increase in about half of the infected cells. These results suggest that taste bud cells after 3 days of culture retain characteristic molecular markers, and may prove useful for describing the molecular and physiological features of taste bud cells, and that these cells can be further manipulated by adenovirus-mediated gene introduction.

Animals↗
PubMed2003

Can the premises of the spasm of resistance vessel concept permit improvement in the treatment and prevention of ischemic heart disease?

In this communication, the spasm of resistance vessel (S-RV) concept of ischemic heart disease (IHD) and other ischemic will be reviewed and updated, and evidence will be presented that principles of the hypothesis might improve the treatment and prevention of IHD. The S-RV concept provides a different basic pathogenetic framework for IHD, and suggestions for treatment and prevention stem from its different basic conceptualization of this disorder. The concept asserts that S-RV directly induces symptoms in IHD, and this position challenges the accepted pathogenetic mechanism for this disorder, i.e., that symptoms in IHD are due directly to obstructive occlusions of epicardial arteries secondary to coronary artery disease. The S-RV concept avers that ischemia-induced S-RV is a major factor in IHD, and evidence supporting this position is provided. Another major position of the hypothesis is that no-reflow (reduced flow after infarction and severe myocardial ischemia in the absence of infarction) is due to ischemic injury-induced S-RV, and a variety of evidences to support this position are offered.Proposed improvement in the treatment of IHD is based mainly on treating ischemia-induced S-RV. alpha-Adrenergic sympathetic blockade reverses ischemia-induced S-RV, and alpha-adrenergic blockade is suggested as therapy for acute coronary syndromes and to prevent complications of percutaneous coronary interventions. Also, angiotensin-converting enzyme inhibition, which has actions similar to alpha-adrenergic blockade, is also suggested. Proposals for the prevention of IHD are based the prevention of S-RV, and special emphasis is given to preventing exercise- and stress-related IHD.

Acute Disease↗
PubMedDate not supplied

A simple modification to permit fast-flow perfusion of brain slices.

An adaptation of a tissue slice chamber is described that stabilizes brain slices during rapid perfusion. Comparison experiments show no deleterious effects of fast perfusion on field potential responses in hippocampal slices, and baseline responses appear to stabilize more quickly with the described modification.

Animals↗
PubMed1991

[The inguinal pedicle flap in emergency traumatic hand surgery: value of the long pedicle permitting an early rehabilitation].

The main drawbacks of the use of a groin pedicle flap in emergency surgery for hand injuries are the declivitous position of the upper limb with a tendency towards oedema and the difficulty of instituting early rehabilitation because of the fact that the injured hand is fixed to the abdomen. Based on an extensive experience of 76 emergency flaps out of more than 100 flaps performed over the last ten years, the authors describe their flap raising technique which allows the formation of a long, closed tube compatible with early rehabilitation and also allowing the possible use of splints.

Emergencies↗
PubMed1996

Cloning of a Xenopus laevis inwardly rectifying K+ channel subunit that permits GIRK1 expression of IKACh currents in oocytes.

Xenopus oocytes injected with GIRK1 mRNA express inwardly rectifying K+ channels resembling IKACh. Yet IKACh, the atrial G protein-regulated ion channel, is a heteromultimer of GIRK1 and CIR. Reasoning that an oocyte protein might be substituting for CIR, we cloned XIR, a CIR homolog endogenously expressed by Xenopus oocytes. Coinjecting XIR and GIRK1 mRNAs produced large, inwardly rectifying K+ currents responsive to m2-muscarinic receptor stimulation. The m2-stimulated currents of oocytes expressing GIRK1 alone decreased 80% after injecting antisense oligonucleotides specific to the 5' untranslated region of XIR, but GIRK1/CIR currents were unaffected. Thus, GIRK1 without XIR or CIR only ineffectively produces currents in oocytes. This result suggests that GIRK1 does not form native homomultimeric channels.

Acetylcholine↗
PubMed1998

Electroencephalographic and histological characteristics of a model of limbic status epilepticus permitting direct control over seizure duration.

Status epilepticus is a neurological emergency associated with substantial morbidity and mortality. Experimental and clinical investigations suggest that prolonged seizure activity is associated with injury to vulnerable neurons. Compounds with neuroprotective properties may minimize such injury. Existing methods of inducing experimental status epilepticus result in seizure activity of variable duration and neuronal injury of variable degree. To minimize such variability, status epilepticus may be stopped with anticonvulsants, but this limits the ability to screen for independent neuroprotective properties. We have developed a simple and reliable non-pharmacological model of limbic status epilepticus in which the duration of status epilepticus is under direct experimental control. Status epilepticus is induced by continuous, unilateral hippocampal stimulation. Using this model, the degree of hippocampal pyramidal cell injury varies in direct proportion to status epilepticus duration across a range of 15-140 min. A progressive sequence of EEG changes unfolds with increasing status epilepticus duration, resembling that seen in other models. This model may serve as a reference against which the effects of potential neuroprotective compounds can be studied.

Animals↗
PubMed2002

A generic strategy for subcloning antibody variable regions from the scFv phage display vector pCANTAB 5 E into pASK85 permits the economical production of F(ab) fragments and leads to improved recombinant immunoglobulin stability.

Apart from the decisive sensitivity and specificity of immunosensors, the employed antibodies essentially contribute to additional key factors like fabrication costs for sensor chips and sensor stability. A production scheme for recombinant antibody fragments has been optimised with respect to these particular issues of biosensor development. The phagemid vector pCANTAB 5 E is widely used for the selection of antibody fragments from corresponding libraries. However, large-scale production of the selected single-chain F(v) (scFv) fragments is substantially restricted by the high cost for the inducer IPTG and the anti-E-tag antibody. The latter is needed in significant amounts for the purification of the recombinant protein. A generic strategy was established for subcloning scFv variable regions from pCANTAB 5 E into the plasmid pASK85 for the expression of F(ab) fragments. pASK85 bears coding sequences for murine constant domains including a His(6) tag at the carboxyl-terminal end of the constant heavy chain domain. The anti-s-triazine antibody K47H served as a model system in this study. Biosynthesis of the F(ab) fragment in a high cell density fermenter was induced by addition of anhydrotetracycline. The F(ab) fragment was subsequently purified from the periplasmic extract in a single step by immobilized metal affinity chromatography (IMAC). A yield of 100 microg/lxOD(550) purified F(ab) fragment was obtained employing a standard fermentation scheme. The sensitivity and cross-reactivity of the F(ab) was comparable to the parent scFv when assayed by enzyme immunoassay. However, the F(ab) fragment exhibited significantly improved long-term stability.

Animals↗
PubMed2002

Telomere looping permits repression "at a distance" in yeast.

In yeast, unlike in higher eukaryotes, transcriptional activators and repressors do not normally work when bound to DNA at large distances (over 500 base pairs) from the gene and, in particular, when positioned downstream of the gene. This restriction is relieved for a transcriptional activator if a gene bearing an activator binding site is placed near a yeast telomere. The explanation proposed is that the folded structure found at the telomere helps appose the DNA-bound activator with proteins binding to the promoter so that recruitment of the transcriptional machinery can be effected "at a distance". Here, we show that a repressor, Tup1, works when tethered to DNA downstream of, and some 1.5-kb from, the gene when the construct is placed near a yeast telomere. The effect, observed with activated as well as basal transcription, is eliminated by deletion of Sir3. These and other results indicate that DNA-tethered Tup1 represses by interacting with some component of the transcriptional machinery binding to the promoter, an interaction that is facilitated by the preformed loop at the telomere.

Base Sequence↗
PubMed1999

Tolerance induction permits the development of graft-versus-host disease: donor-mediated attack following small bowel transplantation in mixed chimeras.

The induction of tolerance to organ allografts would eliminate acute and chronic rejection as well as the need for nonspecific immunosuppression. We have shown that tolerance induced through the creation of mixed allogeneic bone marrow chimeras allows for the long-term engraftment of cardiac and small bowel allografts across strong multiple major histocompatibility barriers. The possibility that tolerance might render the host susceptible to graft-versus-host disease (GVHD) has not been investigated in this or other models of tolerance. To test this possibility chimeras were created by transplantation of T-cell depleted ACI and Lewis bone marrow into lethally irradiated Lewis rats. Chimerism was determined post bone marrow transplant (BMTx) by flow cytometry of lymphocytes from reconstituted animals. ACI/Lew chimeras (ALC), Lewis/ACI F1 (LACF1), and Lewis (LEW) rats all received heterotopic ACI vascularized small bowel grafts. A second group of chimeras received small bowel grafts from ACI rats pretreated with low dose irradiation to eliminate T-cells from the graft. LEW-->LEW small bowel isografts were also performed. Animals were examined for evidence of GVHD by clinical signs and histologic examination of biopsied tissues. GVHD was quantified using the popliteal lymph node enlargement assay. All LACF1 rats developed severe lethal GVHD following ACI small bowel transplant. Bone marrow chimeras, ALC (n = 6), developed fatal GVHD in a similar fashion after receiving a small bowel transplant. LEW-->LEW isografts and chimeras receiving bowel from irradiated ACI rats survived long term without GVHD while ACI-->LEW allogeneic transplants all underwent acute rejection. GVHD or its absence was confirmed histologically. Popliteal lymph node enlargement indices reflected the presence of GVHD in the chimeras (1.87) and LACF1 (5.4) receiving allografts, but not in isografts or chimeras receiving irradiated allogeneic transplants. Analysis of cytokines in the tongues of rats undergoing GVHD showed elaboration of Th1 type proinflammatory cytokines which was not seen in isografted rats or rats receiving preirradiated small bowel. These results demonstrate that tolerance induction through mixed chimerism results in susceptibility to small bowel induced GVHD. Preirradiating the donor bowel prior to SBTx can prevent GVHD.

Animals↗
PubMed2003

A static, closed and scaffold-free bioreactor system that permits chondrogenesis in vitro.

OBJECTIVE: To characterise in vitro engineered cartilaginous constructs made employing a novel static, scaffold-free and closed chamber system. DESIGN: Chondrocytes derived from slaughter age pigs (3-6 months) were seeded at high density (20 x 10(6)) into cylindrical chambers (1.0 x 0.5cm) and were maintained between an upper and a lower membrane (100 kDa) for 21 days and subsequently cultured in open culture for 7 additional days. RESULTS: Viable constructs produced were approximately 10 mmx2mm in size and were stable enough to be handled by surgical pincers. Histology and electron microscopy evaluations revealed a neo-cartilage structure of high cell density with a comprehensive extracellular matrix. Predominately collagen type II and negligible amounts of collagen types I and X were detected using RT-PCR and SDS-PAGE analyses. CONCLUSIONS: In this study, we provide evidence of a scaffold-free system that can produce immature hyaline-like cartilaginous constructs suitable for in vivo implantation, or that may be useful for in vitro studies of events related to the process of chondrogenesis.

Animals↗