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At least 955 records · Page 53Linked to original sources
PubMed2006

Development of a virus-induced gene-silencing system for functional analysis of the RPS2-dependent resistance signalling pathways in Arabidopsis.

Virus-induced gene silencing (VIGS) offers a rapid and high throughput technique platform for the analysis of gene function in plants. Although routinely used in some Solanaceous species, VIGS system has not been well established in Arabidopsis thaliana (L.) Heynh. We have recently reported some factors that potentially influence tobacco rattle virus (TRV)-mediated VIGS of phytoene desaturase (PDS) and actin gene expression in Arabidopsis. In this study, we have further established that the Agrobacterium strain used for agro-inoculation significantly affects the VIGS efficiency. Strain GV3101 was highly effective; C58C1 and LBA4404 were invalid, while EHA105 was plant growth stage-dependent for TRV-induced gene silencing. Furthermore, the VIGS procedure optimised for the PDS gene was applied for the functional analysis of the disease resistance gene RPS2-mediated resistance pathway. Silencing of RPS2 led to loss of resistance to the otherwise avirulence strain of Pseudomonas syringae pv. tomato DC3000 carrying the avirulence gene AvrRpt2. Silencing of RIN4, a RPS2 repressor gene, gave rise to conversion of compatible interaction to incompatible. Silencing of NDR1, RAR1 and HSP90, known to be required for the RPS2-mediated resistance, resulted in loss of the resistance, while silencing of EDS1 and SGT1b, which are not required for the RPS2-mediated resistance, caused no change of the resistance. These results indicate that the optimised procedure for the TRV-based VIGS is a potentially powerful tool for dissecting the signal transduction pathways of disease resistance in Arabidopsis.

Arabidopsis↗
PubMed2005

Recent experiences with minimally invasive pectus excavatum repair "Nuss procedure".

OBJECTIVE: To review the new technical modifications and results of 668 patients who have had pectus excavatum repair utilizing the minimally invasive technique. METHODS: A retrospective chart review was conducted of 668 patients undergoing minimally invasive pectus repair from 1987 through July 2004. Since 1997, a standardized treatment pathway was implemented. Preoperative evaluation included computed tomography (CT) scan, pulmonary function tests, and cardiac evaluations with electrocardiogram and echocardiogram. Indications for operation included at least 2 of the following: progression of the deformity, Haller CT index greater than 3.25, mitral valve prolapse, cardiac compression or displacement, pulmonary function studies that indicate restrictive or obstructive airway disease, previous failed open or minimally invasive pectus repair. Technical and design modifications since 1998 have included routine thoracoscopy, the use of an introducer/dissector for creating the substernal tunnel, elevating the sternum, and routine use of a wired lateral stabilizer and polydioxanone suture (PDS) sutures around the bar and underlying rib to prevent bar displacement. The bar is removed as an outpatient procedure in 2 to 4 years. RESULTS: In 668 patients undergoing minimally invasive pectus repairs, single bars were used in 78.1% and double in 21.7%. Lateral stabilizers were applied in 99.8% and were wired for further stability in 71%. Bar shifts before the use of stabilizers were 14.3%, which decreased to 4.6% after stabilizers were placed and 0.8% with a wired stabilizer and PDS sutures. Results were excellent in 78.5%, good in 13.1%, fair in 4.7% and failed in 3.7% after more than 1 year post bar removal. CONCLUSION: The minimally invasive technique has evolved into an effective method of pectus excavatum repair. Modifications of the technique have reduced complications. Long-term results continue to be excellent.

Adolescent↗
PubMed1984

Effects of physical configuration and chemical structure of suture materials on bacterial adhesion. A possible link to wound infection.

The purpose of this study was to examine the effects of physical configuration and the chemical nature of suture materials on the preferential adherence of bacteria. Ten suture materials of 2-0 (chromic catgut, Dexon, Vicryl, PDS, Mersilene, Tycron, Ethibond, Surgilon, Ethilon, and Prolene) were used. The bacterial strains tested were Staph. aureus and E. coli. The level of bacterial adherence was determined quantitatively by radiolabelled cells and qualitatively by scanning electron microscopy. It was found that the amount of adhered bacteria depended on the type of suture material, the type of bacteria, and the duration of contact. In the group of absorbable sutures, the new PDS sutures exhibited the smallest affinity toward the adherence of both E. coli and Staph. aureus. Dexon sutures had the highest affinity toward these two bacteria. With nonabsorbable sutures, the physical configuration of the sutures contributed more to their ability to attract bacteria than the surface finish. The bacterial adherence on suture materials was also time dependent. Scanning electron microscope morphologic observation also indicated that Staph. aureus adhered on the suture surface in clusters whereas E. coli tended to adhere individually.

Absorption↗
PubMed1985

Isolation and characterization of cartilage proteoglycans immunoreactive with an antibody to skin proteodermatan sulfate core protein.

Low density proteoglycans showing cross-reaction with an antibody to skin proteodermatan sulfate (PDS) core protein were isolated from the bovine articular cartilage, by CsC1 density gradient centrifugation followed by repeated DEAE-cellulose chromatography. The size and amino acid composition of the core proteins of the immunoreactive proteoglycans, eluted at 0.25M and 0.5M NaC1 on DEAE-cellulose column, were quite similar to that of PDS. The glycosaminoglycan components of both proteoglycans were shown to be composed of a hybrid structure of chondroitin sulfate and dermatan sulfate, based on chondroitinase treatments followed by two-dimensional electrophoresis.

Amino Acids↗
PubMed1990

Stimulation of platelet-derived growth factor-induced DNA synthesis by angiotensin II in rabbit vascular smooth muscle cells.

In cultured rabbit vascular smooth muscle cells (VSMCs), angiotensin II by itself had little mitogenic effect even in the presence of cell-free plasma-derived serum (PDS), but markedly stimulated the platelet-derived growth factor (PDGF)-induced DNA synthesis in the presence of PDS. The maximal extent of DNA synthesis induced by PDGF plus angiotensin II was about twice that induced by PDGF alone. The stimulatory effect of angiotensin II was dose-dependent with the maximal response seen at 1 microM and was inhibited by the specific angiotensin II receptor antagonist, [Sar1, Ile8]angiotensin II. In VSMCs, both PDGF and angiotensin II induced expression of the c-fos gene in dose-dependent manners. In contrast to the synergistic effect of angiotensin II and PDGF on DNA synthesis, they induced expression of the c-fos gene in an additive manner. These results suggest that angiotensin II may act as a growth regulator for VSMCs in addition to acting as a vasoconstrictor.

Angiotensin II↗
PubMed1979

Neocortical pyramidal cells: a model with dendritic calcium conductance reproduces repetitive firing and epileptic behavior.

A computer model of a neocortical pyramidal cell has been constructed using ideas similar to those used for hippocampal pyramidal cells. This model has been applied to the study of (a) repetitive firing, and (b) the paroxysmal depolarizing shift (PDS), an important intracellular event during seizures. Although calcium spikes have not been demonstrated directly in neocortical cells, we have postulated (by analogy with hippocampal pyramidal cells) a dendritic calcium conductance and a 'slow potassium' conductance modulated by intracellular calcium ion. With these dendritic ionic conductances, the model is able to reproduce the following experimental features of neocortical pyramidal cells: the afterdepolarization and succeeding afterhyperpolarization after an antidromic spike, and the f-I (firing rate-injected current) curve. Some of the differences between 'fast' and 'slow' pyramidal tract neurons (PTNs) -- narrower spikes and a steeper f-I curve in the fast PTNs -- may be explained by differences in Hodgkin-Huxley potassium kinetics between the two kinds of cell. The same model which faithfully reproduces repetitive firing behavior also reproduces (given appropriate synaptic inputs) the following intracellular events recording during epileptic seizures: (a) a burst of action potentials superimposed on and followed by a PDS, and (b) rapid repetitive firing succeeded by an IPSP. Thus, a single set of parameters can reporduce both normal physiological behavior and 'epileptic' behavior: the particular behavior seen depending on how the cell is stimulated. This overall result is the same as for our model of the CA1 hippocampal cell. It suggests that certain acutely acting epileptogenic agents, e.g. penicillin, may act by increasing synaptic input (perhaps both excitatory and inhibitory) to pyramidal cells, rather than by altering their membrane properties. As in our CA1 hippocampal cell model, bursting seems to be a phenomenon generated by the apical dendrite.

Biophysical Phenomena↗
PubMed1984

The effect of pentylenetetrazol on inward currents of non-bursting neurons and its role in plateau formation.

The epileptogenic drug, pentylenetetrazol (PTZ) produces paroxysmal depolarization shifts in molluscan neurons that are similar to PDSs seen at a mammalian epileptic focus. Most research on molluscan neurons indicates that PTZ acts by altering ionic somatic conductances. This study was carried out to investigate the effect of PTZ on inward currents in isolated neurons of the pond snail, Lymnaea stagnalis, and to investigate how these altered currents might lead to the production of PDSs. In concentrations from 10 to 60 mM, PTZ decreased maximum inward current conductance and shifted the inactivation and activation curves to the left with the former shift being consistently greater. There was no change in reversal potential or time constants for activation and inactivation of inward currents. The effects of the PTZ-induced alterations in the inward currents were studied by incorporating them along with alterations of outward currents seen in this and other studies in a computer model for molluscan neuronal firing. The composite model reproduced in large part the intermediate changes in electrical activity seen before the development of the PDS as well as the PDS.

Animals↗
PubMedDate not supplied

The effects of changing axis II diagnostic criteria.

This study examined whether changes in diagnostic criteria from the DSM-III to the DSM-III for personality disorders (PDs) had the intended effects. Seventy-two subjects at the University of Iowa from three research studies and one clinical sample were administered two structured interviews (the Structured Interview for DSM-III Personality [SIDP] and the revised SIDP [SIDP-R]) to assess DSM-III and DSM-III-R criteria. Major changes in rates of diagnoses were observed between the DSM-III and DSM-III-R criteria with kappas for agreement ranging between -.025 and .571. As expected, the switch from monothetic to polythetic definitions had an effect on which patients were assigned a given diagnosis. However, not all of the other revisions associated with the DSM-III-R had the intended effects. For instance, the frequency of the diagnosis of schizoid PD did not increase, nor did the overlap between borderline and histrionic PDs decrease. In addition, there was an unintended increase in the rate of paranoid PD. An analysis of individual criteria showed how small, apparently minor changes in the wording of criteria can sometimes have major effects on which patients received a diagnosis of PD.

Adult↗
PubMedDate not supplied

Empirical determination of thresholds for case identification: validation of the Personality Diagnostic Questionnaire-Revised.

The Personality Diagnostic Questionnaire-Revised (PDQ-R) was sent to first-degree relatives of major psychotic patients for identification of DSM-III-R personality disorders (PDs). Responses to the PDQ-R were interpreted both literally and empirically, and compared with the Structured Interview for DSM-III PDs (SIDP) as the standard. For literal interpretation, symptoms reported were counted directly for case identification using fixed DSM-III-R thresholds. The empirical approach adjusted the threshold for case identification to maximize concordance with the SIDP. Comparison of the two methods showed that using empirically determined thresholds in some scales gives better concordance with the SIDP. For the dependent and histrionic PD scales, the improvements were statistically significant. The area under the receiver operating characteristic (ROC) curve was computed for each PDQ-R scale to summarize its discriminatory capability across all thresholds. Areas under the ROC curve indicated that the schizoid, schizotypal, borderline, dependent, passive-aggressive, and histrionic PD scales in the PDQ-R have better discriminatory qualities than other PDQ-R scales.

Adult↗
PubMedDate not supplied

Personality disorders do not influence the results of cognitive behavior therapies for anxiety disorders.

Two studies are presented in which the presence of personality disorders (PDs) was assessed using a structured clinical interview (Structured Clinical Interview for DSM-III-R Personality Disorders [SCID-II] by an independent rater before the start of treatment. The therapy effect was measured by change scores. In the first study, patients (N = 31) with panic disorder (with or without agoraphobia) received standardized cognitive behavioral treatment protocols. The effect of treatment as assessed by questionnaires and panic frequency was comparable in the groups with and without PD. In a second study, patients (N = 57) with various anxiety disorders were assessed before and after an individually tailored cognitive behavioral treatment. In general, results showed that anxiety patients with PDs have more severe axis I pathology, but show a change parallel to the patients without PD. Thus, if the effect of therapy is measured by change scores, PD is not related to therapeutic failure of cognitive behavioral treatment of anxiety disorders in general and panic disorder in particular.

Adult↗
PubMed1983

Penicillin-induced epileptic foci in the motor cortex: vertical inhibition.

Neuronal mechanisms responsible for a vertical restriction of focal seizure activity in the motor cortex were analysed. For this purpose intracellular recording from neurones in superficial (50-300 microns below cortical surface), middle (300-800 microns) and deep cortical layers (800-1300 microns) was performed. As a model of foci of various vertical extensions the spread of seizure activity from superficial to deeper cortical laminae following epicortical penicillin application was used. The appearance of characteristic epileptiform potentials in the surface record with a focus restricted to upper cortical laminae was accompanied (i) in superficial neurones by the development of paroxysmal depolarization shifts (PDS), (ii) in middle neurones by depolarization often followed by hyperpolarization, and (iii) in deep neurones by a sequence of membrane potential changes. The latter consisted of an initial depolarization, an early hyperpolarization, an intermediate depolarization, a late hyperpolarization and a final depolarization. The hyperpolarizing components led to complete suppression of action potentials (vertical inhibition). The early hyperpolarization and the first part of the late hyperpolarization were reduced in amplitude when the intracellular chloride activity was elevated. The intermediate depolarization was replaced by PDS with the enlargement of the epileptic focus into the cortex. The actual effect of the vertical inhibition may in part be responsible for the variability in epileptic motor phenomena coinciding with epileptiform potentials in the surface EEG.

Animals↗
PubMed1987

The in vitro lifespan of MRC-5 cells is shortened by 5-azacytidine-induced demethylation.

The minor base 5-methylcytosine (5mC) in DNA may be important for the regulation of gene expression. Random loss of 5mC may occur during pre-replicative DNA synthesis in mortal cell strains, and thus give rise to biochemical aberrations in aging cells. 5-Azacytidine (5azaC) was used to induce loss of 5mC in DNA of human diploid fibroblasts (MRC-5) in an attempt to accelerate in vitro senescence. The 5mC content of DNA was measured by incorporation of [3H]uridine into dividing cells, hydrolysis of DNA and separation of bases by HPLC. In untreated MRC-5 cells, 5mC was 3.6% of the total cytosine (C+5mC) at population doubling (PD) 20 (28% of lifespan) and fell to 1.6% at PD 67 (97% of lifespan). A single pulse treatment with 5azaC (1 microgram/ml) induced demethylation and shortened the lifespan by 10% (6.8 PDs loss). Pulse-treated cells showed temporary growth inhibition, though they subsequently regained normal growth rate and morphology. However, uniform treatment with 0.1 microgram/ml 5azaC between PD 20 and 23 produced no immediate growth inhibition, but a 22% loss of 5mC and 25% decrement in lifespan (16.6 PDs loss). The present results indicate that 5mC levels fall during normal aging of MRC-5 cells and accelerated 5mC loss shortens the in vitro lifespan of these cells. Hypomethylation may thus be responsible for some aspects of in vitro aging.

5-Methylcytosine↗
PubMed1988

Importance of sulfhydryl group for rabbit gastric lipase activity.

We have shown recently that rabbit gastric lipase (RGL) purified from gastric tissue presents catalytic properties comparable with those of human gastric lipase (HGL). We report here that only one sulfhydryl group was modified per molecule of native RGL after incubation at pH 8.0 with 5,5'-dithiobis(2-nitrobenzoic acid) (NbS2) for 4 h or 4,4'-dithiopyridine (4-PDS) for 60 min. With both reagents, a direct correlation was found between the modification of one sulfhydryl group per enzyme molecule and loss of RGL activity. Incubation of RGL with the new hydrophobic sulfhydryl reagent, dodecyldithio-5-(2-nitrobenzoic acid) (C12-NbS), at 30-fold molar excess, at pH 3.0, 5.0 and 8.0, induced immediate and complete inactivation of RGL. Unlike NbS2 and 4-PDS, C12-NbS almost instantaneously stopped the course of tributyrin hydrolysis by RGL, in contrast to porcine pancreatic lipase (PPL). RGL can be included with HGL in the group of sulfhydryl enzymes.

Animals↗
PubMed1989

A preliminary comparison of flat affect schizophrenics and brain-damaged patients on measures of affective processing.

Flat affect is a major component of schizophrenia and is often also seen in neurological disorders. A preliminary set of comparisons were conducted to delineate neuropsychological mechanisms underlying flat affect in schizophrenia, and new measures are described for the assessment of affective deficits in clinical populations. Subjects were schizophrenic with flat affect (SZs), right brain-damaged (RBD), Parkinson's Disease (PDs), and normal control (NC) right-handed adults. Subjects were administered affective measures of expression and perception in both facial and vocal channels. For both perceptual and expressive tasks the SZs performed significantly less accurately than the NCs and the PDs but did not differ from the RBDs. This was the case for both face and voice. This finding lends support to the speculation that right hemisphere mechanisms, especially cortical ones, may be compromised among schizophrenics with flat affect.

Adult↗
PubMed1991

Assessing the affective component of chronic pain: development of the Pain Discomfort Scale.

Few methods exist to assess the affective or reactive dimension of chronic pain, and there are psychometric and practical limitations on the methods that do exist. The current paper reports on the development and validation of the Pain Discomfort Scale, a 10-item instrument designed to fill the need for a brief and psychometrically sound measure of pain affect. Preliminary evidence supports the reliability and validity of the measure. Its internal consistency and test-retest stability coefficients are high. In addition, the results of both correlational and factor analyses of the PDS with other measures support its distinctiveness (from measures of pain intensity) and construct validity (as indicated by its close association with other measures of pain affect). These results support the use of the PDS in situations where a measure of the affective response to chronic pain is needed.

Affect↗
PubMed1995

Laser tissue welding in genitourinary reconstructive surgery: assessment of optimal suture materials.

OBJECTIVES: Laser tissue welding in genitourinary reconstructive surgery has been shown in animal models to decrease operative time, improve healing, and decrease postoperative fistula formation when compared with conventional suture controls. Although the absence of suture material is the ultimate goal, this has not been shown to be practical with current technology for larger repairs. Therefore, suture-assisted laser tissue welding will likely be performed. This study sought to determine the optimal suture to be used during laser welding. METHODS: The integrity of various organic and synthetic sutures exposed to laser irradiation were analyzed. Sutures studied included gut, clear Vicryl, clear polydioxanone suture (PDS), and violet PDS. Sutures were irradiated with a potassium titanyl phosphate (KTP)-532 laser or an 808-nm diode laser with and without the addition of a light-absorbing chromophore (fluorescein or indocyanine green, respectively). A remote temperature-sensing device obtained real-time surface temperatures during lasing. The average temperature, time, and total energy at break point were recorded. RESULTS: Overall, gut suture achieved significantly higher temperatures and withstood higher average energy delivery at break point with both the KTP-532 and the 808-nm diode lasers compared with all other groups (P < 0.05). Both chromophore-treated groups had higher average temperatures at break point combined with lower average energy. The break-point temperature for all groups other than gut occurred at 91 degrees C or less. The optimal temperature range for tissue welding appears to be between 60 degrees and 80 degrees C. CONCLUSIONS: Gut suture offers the greatest margin of error for KTP and 808-nm diode laser welding with or without the use of a chromophore.

Evaluation Studies as Topic↗
PubMed1989

Rat hippocampal slices 'in vitro' display spontaneous epileptiform activity following long-term organotypic culture.

Organotypic cultures of rat hippocampal slices were maintained for periods of up to 12 weeks in vitro. Cultures adopted a two-dimensional architecture whilst retaining the subfields characteristic of intact hippocampal slices. Coventional intracellular onset of spontaneous long-lasting epileptiform activity. Epileptiform activity characteristic of both interictal and ictal events (paroxysmal depolarising shifts, tonic/clonic phases and afterdischarges) was observed in the absence of pharmacological manipulation or of orthodromic stimulation. Epileptiform activity was abolished in the presence of high Mg2+ concentration or tetrodotoxin, agents known to block synaptic transmission. In addition, the frequency of epileptiform events was independent of membrane potential and the amplitude of the paroxysmal depolarising shift (PDS) displayed a near linear relationship with membrane potential. The PDS could be reversed at potentials approaching synaptic equilibrium potential. The N-methyl-D-aspartate (NMDA)-receptor antagonist DL-2-amino-5-phosphonovalerate (DL-APV) dose-dependently reduced both the amplitude and duration of the spontaneous paroxysmal shift, having no effect on the initiation of the event or the resting membrane parameters of the neurone. DL-APV also attenuated a late component of the synaptically evoked excitatory postsynaptic potentials (epsp) not observed in non-epileptiform neurones. Application of GABAA receptor antagonists bicuculline or picrotoxin converted interictal events to ictus. In the presence of these agents, ictal events were up to 90 s in duration. These results suggest that long-term culturing of hippocampal explants leads to an alteration in the balance of excitatory and inhibitory synaptic activity. This allows the expression of an excitatory amino acid depolarisation acting through NMDA receptors which contributes to the generation and maintenance of spontaneous epileptiform activity which is synaptic in origin.

2-Amino-5-phosphonovalerate↗
PubMed1992

A burst-dependent hippocampal excitability defect elicited by potassium at the developmental onset of spike-wave seizures in the Tottering mutant.

Hippocampal CA3 pyramidal neurons in the adult epileptic mutant mouse tottering (tg) show normal intrinsic membrane properties, yet fire abnormally prolonged paroxysmal depolarizing shifts (PDS) during in vitro exposure to elevated extracellular potassium solutions. Intracellular recordings in immature mutants reveal that this network burst abnormality is present during the developmental period that coincides with the onset of seizures in the mutant (19-20 postnatal days), and is significantly more pronounced at this age than at adulthood. These data are inconsistent with the hypothesis that the mutant PDS prolongation represents a secondary consequence of a prolonged history of repeated seizures and suggest that it may reflect a cellular epileptogenic phenotype more directly related to the primary neuropathological expression of the tg gene.

Aging↗