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At least 415 records · Page 23Linked to original sources
PubMed1989

Topical ocular anesthetics affect epithelial cytoskeletal proteins of wounded cornea.

The purpose of this study was to characterize by immunofluorescent microscopy, the cytoskeletal proteins actin and myosin and the protein calmodulin (CaM) in trephined, n-heptanol treated murine corneal epithelium during in vitro organ culture before and following topical ocular anesthetic application. The study has shown that corneal epithelial wounds exposed to anesthetic agents fail to close, even after 24 hr of culture. Failure of the wound to close was accompanied by a general decrease in immunofluorescence for actin myosin and CaM following application of the anesthetics.

Actins↗
PubMed1999

Use of peripheral blood blasts vs bone marrow blasts for diagnosis of acute leukemia.

Acute leukemia can be diagnosed when blasts constitute 30% or more of the nucleated cells in a patient's peripheral blood (PB) sample. To determine whether in such cases bone marrow (BM) aspirates are still necessary, we compared the results of diagnostic studies performed on PB samples with blast counts of 30% or more with those performed on the same patients' BM samples. We found no differences in morphologic features, cytochemistry, or immunophenotype between the blasts in PB and BM samples in any of 30 cases studied. However, in 10 (23%) of 44 cases in which cytogenetic analysis was performed, PB but not BM samples were insufficient for analysis. The converse never occurred. Five of the 10 cases had acute lymphoblastic leukemia and 5 had acute myeloid leukemia (41% of the patients with acute lymphoblastic leukemia and 17% of the patients with acute myeloid leukemia). In cases with adequate metaphases, there was strong correlation between the cytogenetic results for PB and BM samples. Some PB samples with blast counts of 30% or more are adequate for diagnosis of acute leukemia, especially when therapy can be delayed until it is known that an adequate number of analyzable metaphases are recovered from the PB samples.

Acute Disease↗
PubMed2002

Syncytiotrophoblast is a barrier to maternal-fetal transmission of herpes simplex virus.

Herpes simplex virus (HSV)-1 has been discovered in placental tissue from spontaneous miscarriages, but reports of transplacental transmission and fetal infection are extremely rare. Previously, we demonstrated that the villous syncytiotrophoblast, which forms a continuous layer between the maternal and fetal circulation, is resistant to HSV entry. Here, we tested our hypothesis that the villous syncytiotrophoblast prevents transplacental transmission of HSV secondary to decreased expression of HSV entry mediators (HveA, HveB, and HveC). In addition, we investigated the ability of HSV to infect extravillous trophoblast cells, which mediate placental attachment to the uterine wall, and the expression of HSV receptors in these cells. We performed fluorescence-activated cell sorting (FACS) analyses and immunostaining to demonstrate that HveA, HveB, and HveC were not expressed in third-trimester villous trophoblast cells. Consequently, villous explants obtained from third-trimester placentas were resistant to infection by a recombinant HSV-1 vector, HSV-1 KOS, but approximately 20% of mesenchymal cells within the villous core were infected when villous explants were pretreated with trypsin to disrupt the villous trophoblast layer. Conversely, FACS analysis and immunostaining demonstrated that extravillous trophoblast cells expressed HveA, HveB, and HveC, and these cells were efficiently infected by HSV vectors. Infection of extravillous trophoblast cells by HSV-1 was not reduced when the cells were pretreated with an antibody against HveA but was partially reduced when the cells were pretreated with antibodies directed against HveB and HveC. Thus, the decreased expression of herpesvirus entry mediators in villous syncytiotrophoblast prevents placental villous infection, thereby limiting maternal-fetal transmission of HSV.

Cell Separation↗
PubMed1999

Analysis of the effect of conversion from open to closed surgical intensive care unit.

OBJECTIVE: To compare the effect on clinical outcome of changing a surgical intensive care unit from an open to a closed unit. DESIGN: The study was carried out at a surgical intensive care unit in a large tertiary care hospital, which was changed on January 1, 1996, from an open unit, where private attending physicians contributed and controlled the care of their patients, to a closed unit, where patients' medical care was provided only by the surgical critical care team (ABS or ABA board-certified intensivists). A retrospective review was undertaken over 6 consecutive months in each system, encompassing 274 patients (125 in the open-unit period, 149 in the closed-unit period). Morbidity and mortality were compared between the two periods, along with length-of-stay (LOS) and number of consults obtained. A set of independent variables was also evaluated, including age, gender, APACHE III scores, the presence of preexisting medical conditions, the use of invasive monitoring (Swan-Ganz catheters, central and arterial lines), and the use of antibiotics, low-dose dopamine (LDD) for renal protection, vasopressors, TPN, and enteral feeding. RESULTS: Mortality (14.4% vs. 6.04%, p = 0.012) and the overall complication rate (55.84% vs. 44.14%, p = 0.002) were higher in the open-unit group versus the closed-unit group, respectively. The number of consults obtained was decreased (0.6 vs. 0.4 per patient, p = 0.036), and the rate of occurrence of renal failure was higher in the open-unit group (12.8% vs. 2.67%, p = 0.001). The mean age of the patients was similar in both groups (66.48 years vs. 66.40, p = 0.96). APACHE III scores were slightly higher in the open-unit group but did not reach statistical significance (39.02 vs. 36.16, p = 0.222). There were more men in the first group (63.2% vs. 51.3%). The use of Swan-Ganz catheters or central and arterial lines were identical, as was the use of antibiotics, TPN, and enteral feedings. The use of LDD was higher in the first group, but the LOS was identical. CONCLUSIONS: Conversion of a tertiary care surgical intensive care unit from an open to closed environment reduced dopamine usage and overall complication and mortality rates. These results support the concept that, when possible, patients in surgical intensive care units should be managed by board-certified intensivists in a closed environment.

Aged↗
PubMedDate not supplied

Evaluation of the harmonized alert sensing technology device for hemodynamic monitoring in chronic hemodialysis patients.

The Harmonized Alert Sensing Technology (HASTE) device was developed to overcome the primary shortcomings of interval based noninvasive blood pressure (BP) monitoring. This study was conducted to assess the reliability of the HASTE system compared with standard cuff BP values in patients on hemodialysis. A total of 1,370 HASTE measurements were compared with oscillometric standard cuff systolic BP values in 42 sessions of 15 patients on hemodialysis. The average discrepancy between the HASTE and cuff systolic BP was 1.41 +/- 16.90 mm Hg. Compared with cuff measurements, 31% of systolic BP fell within a range of 5 mm Hg difference, 57% of systolic BP fell within 10 mm Hg, and 73% of systolic BP fell within a 15 mm Hg band. According to British Hypertension Society standards or Association for the Advancement of Medical Instrumentation criteria, the current HASTE method did not perform well. Technology to provide noninvasive hemodynamic monitoring is, however, in its developmental stage. The effort at continuous systolic pressure monitoring using existing, readily available, and frequently used techniques is exciting. Although the HASTE system as currently configured and calibrated did not adequately perform, variations in site analysis and conversion factors may increase pressure sensitivity and tracking over the course of a standard dialysis treatment.

Adult↗
PubMed1990

Short-term variability of systolic blood pressure and heart rate in normotensive subjects.

Short-term fluctuations in systolic blood pressure (SBP) and heart rate (HR) and their inter-relationship were analysed in a group of normotensive middle-aged men (n = 16) using a multivariate autoregressive modelling technique. This study is the first to evaluate the beat-to-beat variability of SBP and HR in a group of real normotensive subjects. Direct intra-arterial blood pressure was registered together with ECG using an ambulatory tape recording technique (the Oxford method). Power spectrum density estimated (PSD) were used as a measure of the variability. PSDs were calculated over 3-min periods for four basic physiological conditions: during sleep and in the supine, sitting and standing positions. The inter-relationship between the blood pressure and heart rate variabilities was analysed using a closed-loop model. In agreement with results presented earlier in the literature, the beat-to-beat variation in SBP and HR was concentrated in three typical power spectrum regions: the high-frequency (HF = 0.15-0.35 Hz) region (respiration), the mid-frequency (MF = 0.075-0.15 Hz) region (vasomotor oscillation) and the low-frequency (LF = 0.02-0.075 Hz) region (thermoregulation). The variability changes considerably between different situations, especially that of the MF region. The variability was most prominent in the MF region and in the standing position. The variability was generally smallest in the HF region and in sleep. The results also demonstrate that the beat-to-beat variability in SBP and HR can considerably affect one another.

Adult↗
PubMed1997

Rhetorical intentionality attribution: its ontogenesis in ordinary conversation.

This paper addresses the assumption that the epistemological dichotomy between reality and its construction is an everyday rhetorical tool children and adults use in their explanations and arguments. Within a discursive psychology perspective, the phenomenon of attribution of rhetorical intentionality (AIR) is theoretically defined developing the notion of prepackaged interest attribution technique (Edwards & Potter, 1992). AIR consists in an explanation of others' discursive moves in terms of a rhetorical-argumentative activity; that is, in terms of a deliberate attempt aimed at construing and presenting reality as a function of the speaker's point of view, perspective and interests, in order to affect the listeners mental state (belief, knowledge, intention) or action. Theoretical links are made to the theory of mind paradigm in the study of intentionality attribution, particularly to the form of life approach which defines intentionality attribution in terms of language games. On this basis, the present study examines the structures and functions of AIR used by children and their parents in everyday conversation. Qualitative discourse analysis of transcripts of 22 dinner conversations from eight Italian families shows explicit and implicit AIR in children from three and a half to seven years of age. Moreover, parents attribute rhetorical intentionality to their children, contributing to their social construction as rhetorical speakers. AIR is used as a rhetorical strategy within disputes and argumentative contexts, often achieving topic closure. Results are discussed with reference to the ontological status of intentionalist talk.

Adult↗
PubMed1997

Nucleotide sequences and regulational analysis of genes involved in conversion of aniline to catechol in Pseudomonas putida UCC22(pTDN1).

A 9,233-bp HindIII fragment of the aromatic amine catabolic plasmid pTDN1, isolated from a derivative of Pseudomonas putida mt-2 (UCC22), confers the ability to degrade aniline on P. putida KT2442. The fragment encodes six open reading frames which are arranged in the same direction. Their 5' upstream region is part of the direct-repeat sequence of pTDN1. Nucleotide sequence of 1.8 kb of the repeat sequence revealed only a single base pair change compared to the known sequence of IS1071 which is involved in the transposition of the chlorobenzoate genes (C. Nakatsu, J. Ng, R. Singh, N. Straus, and C. Wyndham, Proc. Natl. Acad. Sci. USA 88:8312-8316, 1991). Four open reading frames encode proteins with considerable homology to proteins found in other aromatic-compound degradation pathways. On the basis of sequence similarity, these genes are proposed to encode the large and small subunits of aniline oxygenase (tdnA1 and tdnA2, respectively), a reductase (tdnB), and a LysR-type regulatory gene (tdnR). The putative large subunit has a conserved [2Fe-2S]R Rieske-type ligand center. Two genes, tdnQ and tdnT, which may be involved in amino group transfer, are localized upstream of the putative oxygenase genes. The tdnQ gene product shares about 30% similarity with glutamine synthetases; however, a pUC-based plasmid carrying tdnQ did not support the growth of an Escherichia coli glnA strain in the absence of glutamine. TdnT possesses domains that are conserved among amidotransferases. The tdnQ, tdnA1, tdnA2, tdnB, and tdnR genes are essential for the conversion of aniline to catechol.

Amino Acid Sequence↗
PubMed1997

A novel 165-base-pair terminal repeat sequence is the sole cis requirement for the adeno-associated virus life cycle.

Adeno-associated virus (AAV) replication is dependent on two copies of a 145-bp inverted terminal repeat (ITR) that flank the AAV genome. This is the primary cis-acting element required for productive infection and the generation of recombinant AAV (rAAV) vectors. We have engineered a plasmid (pDD-2) containing only 165 bp of AAV sequence: two copies of the D element, a unique sequence adjacent to the AAV nicking site, flanking a single ITR. When assayed in vivo, this modified hairpin was sufficient for the replication of the plasmid vector when Rep and adenovirus (Ad) helper functions were supplied in trans. pDD-2 replication intermediates were characteristic of the AAV replication scheme in which linear monomer, dimer, and other higher-molecular-weight replicative intermediates are generated. Compared to infectious AAV clones for replication, the modified hairpin vector replicated more efficiently independent of size. Further analysis demonstrated conversion of the input circular plasmid to a linear substrate with AAV terminal repeat elements at either end as an initial step for replication. This conversion was independent of both Rep and Ad helper genes, suggesting the role of host factors in the production of these molecules. The generation of these substrates suggested resolution of the modified terminal repeat through a Holliday-like structure rather than replication as a mechanism for rescue. Production of replicative intermediates via this plasmid substrate were competent not only for AAV DNA replication but also for encapsidation, infection, integration, and subsequent rescue from the chromosome when superinfected with Ad and wild-type AAV. These studies demonstrate that this novel 165-bp ITR substrate is sufficient in cis for the AAV life cycle and should provide a valuable reagent for further dissecting the cis sequences involved in AAV replication, packaging, and integration. In addition, this novel plasmid vector can be used as a substrate for both rAAV vector production and synthetic plasmid vector delivery.

DNA, Viral↗
PubMed1985

Structure and transcription of a telomeric surface antigen gene of Trypanosoma brucei.

The gene encoding variant surface glycoprotein 221 in Trypanosoma brucei is located adjacent to a chromosome end and can be activated with or without a concomitant gene duplication. To test whether transcription initiates within the cloned segment of the 221 gene, we analyzed nascent and stable transcripts. We show here that the 221 coding region and 8.5 kilobases of adjacent upstream DNA are transcribed into nascent RNA at a similar rate when gene 221 is activated without duplication. Since only part of this transcribed upstream segment is transferred with the coding region to another telomere upon duplicative activation of gene 221, we infer that initiation of variant surface glycoprotein gene transcription occurs outside the gene segment that moves into an expression site by gene conversion. Our analysis shows that part of the variant surface glycoprotein 221 transcription unit consists of an unusual 3.5-kilobase tandem array of ca. 50 repeat segments and that a rearrangement in this array accompanies the nonduplicative activation of gene 221. A variant surface glycoprotein pseudogene is located within the transcription unit of gene 221, and we discuss models that account for this unusual situation.

Animals↗
PubMed2000

The influence of elbow joint angle on different phases of force development during maximal voluntary contraction.

The first aim of the study was to find an elbow joint angle at which muscle can produce maximum voluntary force (Lo(MVC)) and to compare that angle with an angle at which the fastest rates of force development occur (Lo). The second aim of the study was to find if changes in MVC and force development speed at an angle smaller (Ls) and larger (Ll) than the optimal angle depend on whether Ls and Ll were compared to Lo or Ls and Ll to Lo(MVC). Twenty-four male physical education students were tested four times using the BIODYNA dynamometer to measure torque versus time at an optimal length, as well as at lengths that were shorter (Ls = optimal -30 degrees) and longer (Ll = optimal +50 degrees). The average values of optimal angles for force development indices (Lo) were similar to the angle at which maximum force was produced (Lo(MVC)); however, there was a small (5-10 degrees) difference between Lo and Lo(MVC) in the majority of subjects. The results showed that during elbow flexion with the forearm in the midrange position, the difference between Lo and Lo(MVC) was small and did not affect MVC; however, it had a significant effect on the relation between joint angle and force development speed.

Adult↗
PubMed2005

Characteristics and actions of NAD(P)H oxidase on the sarcoplasmic reticulum of coronary artery smooth muscle.

It has been reported that nonmitochondrial NAD(P)H oxidases make an important contribution to intracellular O2-* in vascular tissues and, thereby, the regulation of vascular function. Topological analyses have suggested that a well-known membrane-associated NAD(P)H oxidase may not release O2-* into the cytosol. It is imperative to clarify the source of intracellular O2-* associated with this enzyme and its physiological significance in vascular cells. The present study hypothesized that an NAD(P)H oxidase on the sarcoplasmic reticulum (SR) in coronary artery smooth muscle (CASM) regulates SR ryanodine receptor (RyR) activity by producing O2-* locally. Western blot analysis was used to detect NAD(P)H oxidase subunits in purified SR from CASM. Fluorescent spectrometric analysis demonstrated that incubation of SR with NADH time dependently produced O2-*, which could be substantially blocked by the specific NAD(P)H oxidase inhibitors diphenylene iodonium and apocynin and by SOD or its mimetic tiron. This SR NAD(P)H oxidase activity was also confirmed by HPLC analysis of conversion of NADH to NAD+. In experiments of lipid bilayer channel reconstitution, addition of NADH to the cis solution significantly increased the activity of RyR/Ca2+ release channels from these SR preparations from CASM, with a maximal increase in channel open probability from 0.0044 +/- 0.0005 to 0.0213 +/- 0.0018; this effect of NADH was markedly blocked in the presence of SOD or tiron or the NAD(P)H oxidase inhibitors diphenylene iodonium, N-vanillylnonanamide, and apocynin. These results suggest that a local NAD(P)H oxidase system on SR from CASM regulates RyR/Ca2+ channel activity and Ca2+ release from SR by producing O2-*.

Animals↗
PubMed1980

Prostaglandin E2 production by rabbit urinary bladder.

Synthesis of prostaglandin E2 (PGE2) by rabbit bladder was examined. PGE2 synthesis was assessed by thin-layer chromatographic analysis after conversion of [14C]-arachidonic acid to [14C]PGE2 or by a specific radioimmunoassay technique. Intact bladder and microsomes prepared from the bladder transitional epithelium (mucosal) layer and the outer vesicular layer demonstrated synthesis of PGE2. PGE2 synthesis was increased by arachidonic acid and blocked by indomethacin. When the inside medium (bathing the transitional epithelium) contained [14C]arachidonic acid, no detectable radioactivity was observed in the outside medium (bathing the outer layer). Conversely, when the outside medium contained [14C]arachidonic acid, no detectable radioactivity was observed in the inside medium. In addition, [14C]arachidonic acid was incorporated only into tissue directly exposed to bathing media containing the label. These results demonstrate that the rabbit bladder can synthesize PGE2 and that the PGE2- synthesizing systems of the transitional epithelium and outer layer of bladder may be distinct.

Animals↗
PubMed1989

Transition from dynamically maintained to relaxed end-expiratory volume in human infants.

Newborn infants, in contrast to adults, dynamically maintain end-expiratory lung volume (EEV) above relaxation volume. The purpose of this study was to determine at what age children develop a breathing strategy that is relaxed, i.e., determined by the mechanical characteristics of the lung and chest wall. Forty studies were performed in 27 healthy infants and children aged 1 mo to 8 yr during natural sleep. Volume changes were recorded with the use of respiratory inductance plethysmography (RIP). The volume signal was differentiated to yield flow. Flow-volume representations were generated for a random sample of the recorded breaths to determine the predominant breathing strategy utilized, i.e., relaxed, interrupted, or indeterminate. The respiratory pattern was predominantly interrupted below 6 mo of age and predominantly relaxed over 1 yr of age. Mixed patterns were observed in children 6-12 mo of age. The number of breaths that could not be classified (indeterminate) decreased with age. Respiratory frequency measured from the sample of breaths decreased with age and was accompanied by an increase in expiratory time. We conclude that a relaxed EEV develops at the end of the first year of life and may be related to changes in the mechanical properties of the chest wall associated with growth as well as changes in respiratory timing.

Analog-Digital Conversion↗
PubMed2005

Limitations of commonly used internal controls for real-time RT-PCR analysis of renal epithelial-mesenchymal cell transition.

BACKGROUND/AIMS: Progressive renal fibrotic disease is accompanied by the massive accumulation of myofibroblasts as defined by alpha smooth muscle actin (alphaSMA) expression. We quantitated gene expression using real-time RT-PCR analysis during conversion of primary cultured human renal tubular cells (RTC) to myofibroblasts after treatment with transforming growth factor-beta1 (TGF-beta1). We report herein the limitations of commonly used reference genes for mRNA quantitation. METHODS: We determined the expression of alphaSMA and megakaryoblastic leukemia-1 (MKL1), a transcriptional regulator of alphaSMA, by quantitative real-time PCR using three common internal controls, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cyclophilin A and 18S rRNA. RESULTS: Expression of GAPDH mRNA and cyclophilin A mRNA, and to a lesser extent, 18S rRNA levels varied over time in culture and with exposure to TGF-beta1. Thus, depending on which reference gene was used, TGF-beta1 appeared to have different effects on expression of MKL1 and alphaSMA. CONCLUSIONS: RTC converting to myofibroblasts in primary culture is a valuable system to study renal fibrosis in humans. However, variability in expression of reference genes with TGF-beta1 treatment illustrates the need to validate mRNA quantitation with multiple reference genes to provide accurate interpretation of fibrosis studies in the absence of a universal internal standard for mRNA expression.

Actins↗
PubMed1990

Depth of occlusal caries assessed clinically, by conventional film radiographs, and by digitized, processed radiographs.

Occlusal caries depth was assessed in 47 extracted premolars and permanent molars by 4 observers on a rank scale by eye inspection, by film radiographs, and by 2 of the observers also by digitized radiographs after filtering and contrast enhancement of the image. Quantitative estimates of caries depth were obtained from the digitized radiographs. Accuracy of scorings was determined with the histologic section as validating criterion. Occlusal caries was present in a spectrum from incipient fissure decalcification to large cavity formation. Clinical as well as radiographic scorings most frequently under-estimated lesion depth. Accuracy of radiographic assessments increased substantially by digital processing of the radiographic image. Quantitative measures of caries depth on digitized radiographs were strongly correlated to the histologic measures (r = 0.91). Interobserver agreement was fair to moderate according to kappa coefficients for the clinical and the radiographic scorings. The agreement was highest for the scorings of the digitally enhanced images. The study suggests that digital processing of radiographic images constitutes a diagnostic aid that may give a more accurate estimate of occlusal caries depth.

Analog-Digital Conversion↗