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At least 1,171 records · Page 65Linked to original sources
PubMed1999

Chronopharmacology for deoxyspergualin: toxicity and efficacy in the rat.

BACKGROUND: Deoxyspergualin (DSG), a potent immunosuppressive drug, has been clinically used as a rescue therapy for acute graft rejection. However, DSG has severe adverse effects that limits its use. Here, we examined the optimized therapeutic protocol for DSG using chronopharmacological profiles of it. METHODS: First, a dosing time-dependent difference in the toxicity and efficacy of DSG was determined using a heterotopic heart transplantation model. Second, chronokinetics of DSG were examined to reveal the mechanism for dosing time-dependent differences in the effect. RESULTS: In rats treated with DSG at the active period, bone marrow suppression and damage of small intestine were significantly severe. However, no significant difference was found in cardiac allograft survival and pharmacokinetics according to the timing of drug administration. CONCLUSIONS: The toxicity of DSG varied with the dosing time, whereas its efficacy did not. The chronopharmacological approach may provide merits for immunosuppressive therapy with DSG in clinical organ transplantation.

Animals↗
PubMed2002

Combination induction therapy with monoclonal antibodies specific for CD80, CD86, and CD154 in nonhuman primate renal transplantation.

BACKGROUND: Antibodies and fusion proteins specific for CD80, CD86, and CD154 have shown promise as agents capable of inducing donor-specific tolerance in rodents. These agents have also been shown to be synergistic with one another in many settings of counter-adaptive immunity. In the nonhuman primate, monoclonal antibodies specific for CD80 and CD86 have prolonged the time to rejection of renal allografts but have not resulted in tolerance. A monoclonal antibody specific for CD154 has resulted in markedly prolonged survival of kidney, islet, cardiac, and skin allografts, but again most animals have eventually developed rejection after prolonged periods of rejection-free survival off therapy. METHODS: A combination of monoclonal antibodies specific for CD80, CD86, and CD154 were used in a mismatched nonhuman primate renal-allograft model. Doses used were based on optimized treatment protocols for each agent individually. RESULTS: Treatment of four rhesus macaques with this combination yielded a mean rejection-free survival of 565 days (311-911 days), significantly greater than untreated controls (mean survival=7.0 days, P=0.001) and animals treated with only a combination of anti-CD80 and CD86 (mean survival=191 days, P=0.01). The survival of animals treated with this combination of monoclonal antibodies was not significantly greater than those treated with anti-CD154 alone, but the production of alloantibody was delayed compared with monotherapy anti-CD154. CONCLUSION: These data suggest that a synergy exists between these agents, particularly with regard to T-dependent B-cell responses, but that they fail to induce durable tolerance in nonhuman primates.

Animals↗
PubMed1995

Epilepsy surgery in adults.

Epilepsy surgery is capable of stopping seizures or markedly reducing their frequency in selected patients with medically refractory epilepsy. Presurgical evaluation, as currently practiced, requires concordance among a battery of localizing tests. Cost-effectiveness is increasingly important in the development of optimal presurgical protocols and surgical procedures in specific patient populations.

Adult↗
PubMed2004

Magnetic resonance imaging of suspected cervicocranial arterial dissections.

The authors propose that the optimal screening protocol for evaluation of suspected cervicocranial arterial dissections is magnetic resonance imaging (MRI) that includes three components: 1) contrast-enhanced three-dimensional time-of-flight magnetic resonance angiography (MRA) through the superior mediastinum, neck, and skull base; 2) three-dimensional multiple overlapping thin-section acquisition MRA of the skull base and Circle of Willis region; and 3) axial non-contrast, non-fat-suppressed T1-weighted, fat-suppressed T1-weighted, and T2-weighted spin-echo MRI from the level of the aortic arch through the level of the circle of Willis. MRA permits visualization of vascular luminal narrowing or obliteration, which can suggest vascular dissection but can also be caused by congenital variation, dysplasia, intraluminal thrombus, vasospasm, or extramural compression by tumor. By directly visualizing the blood vessel wall, axial T1-weighted and T2-weighted spin-echo MRI can identify the intramural hemorrhage of vascular dissection. This protocol is designed to maximize the sensitivity of a noninvasive technique and may eliminate the need for conventional endovascular angiography.

Adult↗
PubMed2004

Prevention of hospital-associated pneumonia and ventilator-associated pneumonia.

OBJECTIVE: To synthesize the available clinical data for the prevention of hospital-associated pneumonia (HAP) and ventilator-associated pneumonia (VAP) into a practical guideline for clinicians. DATA SOURCE: A Medline database and references from identified articles were used to perform a literature search relating to the prevention of HAP/VAP. CONCLUSIONS: There is convincing evidence to suggest that specific interventions can be employed to prevent HAP/VAP. The evidence-based interventions focus on the prevention of aerodigestive tract colonization (avoidance of unnecessary antibiotics and stress ulcer prophylaxis, use of sucralfate for stress ulcer prophylaxis, chlorhexidine oral rinse, selective digestive decontamination, short-course parenteral prophylactic antibiotics in high-risk patients) and the prevention of aspiration of contaminated secretions (preferred oral intubation, appropriate intensive care unit staffing, avoidance of tracheal intubation with the use of mask ventilation, application of weaning protocols and optimal use of sedation to shorten the duration of mechanical ventilation, semirecumbent positioning, minimization of gastric distension, subglottic suctioning, avoidance of ventilator circuit changes/manipulation, routine drainage of ventilator circuit condensate). Clinicians caring for patients at risk for HAP/VAP should promote the development and application of local programs encompassing these interventions based on local resource availability, occurrence rates of HAP/VAP, and the prevalence of infection due to antibiotic-resistant bacteria (Pseudomonas aeruginosa, Acinetobacter species, and methicillin-resistant Staphylococcus aureus).

Cross Infection↗
PubMed2005

Molecular strategies for identifying human papillomavirus infection in routinely processed samples: focus on paraffin sections.

OBJECTIVE: To optimize the protocols of human papillomavirus (HPV) detection in clinical samples, we used polymerase chain reaction (PCR)-based techniques in paraffin-embedded tissue sections and compared the results with those obtained with PCR and Hybrid Capture II (HC2) performed in liquid-based cytology (LBC) preservation medium. MATERIALS AND METHODS: Forty-five consecutive cervical biopsy specimens were taken from women participating in the ongoing Latin American Screening Study at Leonor Mendes de Barros Hospital, São Paulo, Brazil, during 2003 and 2004. The biopsy specimens were analyzed for HPV by a modified GP5+/GP6+ PCR protocol, and the results were compared with those obtained by PGMY PCR and HC2 in samples collected in LBC preservation medium. RESULTS: beta-Globin was detected in 100% of the multiplex PCR system from LBC samples and 66.7% with PCO4+/PCO3+ PCR in biopsy specimens. Of the three methods, PGMY PCR system and HC2 were equally effective in detecting HPV; both detected 13 cases in 45 samples (28.9%). The GP5+/GP6+ PCR applied in biopsy specimens showed a 20% HPV detection rate (9/45). CONCLUSIONS: Our PCR protocols worked reasonably well and allowed us to compare the three molecular methods with histological and cytological findings. The reproducibility of the results makes the technique applicable in archival materials.

Adolescent↗
PubMed2004

Villous adenocarcinoma of the colon and rectum: a clinicopathologic study of 36 cases.

Some colorectal adenocarcinomas show villous architecture with morphologic similarities to tubulovillous or villous adenomas. We reviewed 420 consecutive colorectal adenocarcinoma resection specimens and found that 95 tumors (23%) showed areas of villous architecture. Thirty-six tumors (8.6%) in 35 patients showed more than 50% villous architecture and were designated villous adenocarcinomas. Only 42% of the villous adenocarcinomas showed severe atypia and only 44% of the available pre-resection biopsies of these tumors were diagnosed as adenocarcinoma. Epithelial islands in desmoplastic stroma (EIDS) may be helpful in the diagnosis of these tumors. EIDS were found in 97% of the resection specimens for villous adenocarcinomas and none of 62 resection specimens for tubulovillous or villous adenomas. The presence of EIDS showed a 67% sensitivity, 100% specificity, and 100% predictive value in the diagnosis of villous adenocarcinoma in a blinded review of villous tumors. On review of the pre-resection biopsies of villous adenocarcinoma without a final diagnosis of adenocarcinoma, 40% showed EIDS. Clinical follow-up of the 35 patients with villous adenocarcinoma showed that only one died of colorectal adenocarcinoma (median follow-up, 46 months). This sole patient dying of colorectal adenocarcinoma showed a synchronous advanced stage of nonvillous adenocarcinoma at the time of diagnosis. Villous adenocarcinoma is a diagnostically challenging subset of colorectal adenocarcinoma, which appears to be associated with a favorable prognosis. Classifying these tumors as a special type of colorectal cancer may facilitate the development of diagnostic adjuncts and optimal treatment protocols.

Adenocarcinoma↗
PubMed2006

Ultrasonic decalcification offers new perspectives for rapid FISH, DNA, and RT-PCR analysis in bone marrow trephines.

The requisite analyses on bone marrow biopsies are increasing: Molecular analyses such as fluorescence in situ hybridization (FISH), polymerase chain reaction (PCR), and reverse transcriptase (RT)-PCR are demanded in addition to morphology and immunohistochemistry to improve diagnostic accuracy. Moreover, analysis of certain molecular prognostic or predictive biomarkers is increasingly mandatory in the assessment of hematologic diseases. In some circumstances, only formalin fixed, bone-containing tissue is available for molecular analysis. Because various fixation and decalcification procedures can impair DNA and RNA quality, there is an urgent need for standardized decalcification protocols which allow FISH and PCR analysis. In this study we developed a routinely applicable decalcification protocol to optimize the molecular analysis method although preserving morphology and immunohistochemical results. Therefore, we compared 2 different approaches including ultrasonic decalcification versus nonultrasonic procedures and ethylenediaminetetraacetate-based reagents versus acid-based ones. In our hands, the combined use of ultrasound and ethylenediaminetetraacetate-based reagents permits successful interphase FISH, PCR, and RT-PCR analysis whereas concomitantly preserving morphology and antigeneicity.

Biopsy↗
PubMed2004

Multislice CT virtual intravascular endoscopy for abdominal aortic aneurysm stent grafts.

PURPOSE: To investigate the optimal scanning protocols of multislice computed tomographic (CT) angiography in aortic stent graft placement observed on virtual intravascular endoscopy (VIE). MATERIALS AND METHODS: A human aorta phantom was built with a commercial aortic stent graft placed in situ. The phantom was housed in a perspex container and filled with contrast medium with CT attenuation similar to that used in the patient's abdominal scanning. CT was performed with a four-slice multislice CT scanner with section thickness of 1.3 mm, 2 mm, and 3.2 mm, pitch of 0.75, 1, and 1.5, and reconstruction interval of 50% overlap and no overlap. Stair-step artifacts were quantified by measuring the SD of signal intensity on surfaced shaded VIE images in three different locations: superior mesenteric artery (SMA), renal ostium, and aortic aneurysm. RESULTS: Image quality of VIE images was independent of section thickness and pitch values in the level of renal ostium and aortic aneurysm (P >.05), whereas it was determined by the section thickness and pitch in the level of SMA (P <.05). Aortic branch origin became irregular or distorted when section thickness increased to 3.2 mm or pitch reached 1.5. CONCLUSION: A scanning protocol of section thickness of 2 mm, pitch 1, and reconstruction interval of 1 mm is recommended in aortic stent graft placement because it allows fewer stair-step artifacts and better visualization of the aortic stent wires observed on VIE.

Aortic Aneurysm, Abdominal↗
PubMed2003

Crush injury and crush syndrome: a review.

Crush injuries and the subsequent systemic manifestations are well described, but there is limited evidence on which to base protocols for optimal management. Given that in most environments such in-juries are relatively rare, there is, however, an undoubted need for such guidelines. This article reviews the currently available evidence and summarizes the treatment options available.

Crush Syndrome↗
PubMed2006

Visceral torso computed tomography for clearance of the thoracolumbar spine in trauma: a review of the literature.

BACKGROUND: Accurate screening of the thoracolumbar spine (TLS) remains problematic in the care of trauma patients. The current standard of care for TLS screening is not clearly defined. In trauma patients undergoing computed tomography (CT) of the chest and or abdomen, compelling supportive evidence for reformatting and reusing this CT data to clear the thoracolumbar spine has accumulated over the last 3 years. The objective of this review was to identify and review all published studies comparing reformatted CT to traditional plain radiography for TLS clearance. METHODS: A Medline search for all English language articles published on this subject since 1980 identified seven studies. Each was classified according to the levels of evidence classification of the Agency for Health Care Policy and Research. The methodology of each study was reviewed for the CT protocol, utilization of radiologist blinding and whether historical dictated reports or de novo image readings were used for comparison. The sensitivity and specificity of each study and the gold standard utilized in its calculation was noted. Where available, detailed information regarding the missed injuries and their clinical relevance was abstracted for each study. RESULTS: All evaluated studies demonstrated superior sensitivity and interobserver variability for reformatted CT compared with plain radiographic screening. CT was also more accurate in localizing, classifying, and delineating the age, bony intrusion, and soft-tissue damage associated with the fracture. For studies with time-motion components, a protocol utilizing CT clearance was not only more accurate but faster and more economical. Screening with reformatted visceral CT data required no additional scan time or radiation exposure. CONCLUSION: The evidence to date demonstrates the superior sensitivity of reformatted visceral CT for detecting thoracolumbar spine injury. With no further patient movement, radiation exposure, cost, or time, trauma patients undergoing visceral CT can have their thoracolumbar spine promptly evaluated. Further prospective evaluation of the CT protocols to optimize visualization of both the viscera and the bone is warranted.

Adult↗
PubMed2004

Noradrenaline use in the human donor and relationship with load-independent right ventricular contractility.

BACKGROUND: Experimental and clinical studies suggest that brain death (BD)-associated cardiac dysfunction is related to the neurohormonal storm and subsequent exposure to intravenous catecholamines. We aimed to describe the relationship between empirical noradrenaline treatment and donor heart function, described for the first time with load-independent indices of right ventricular contractility. METHODS: Twenty-seven BD patients were divided in three groups based on noradrenaline at time of offer, started by the donor hospital: group 1=no noradrenaline (n=11); group 2=low dose (n=8); group 3=high dose (n=8). After protocol-guided optimization by our retrieval team using a Swan-Ganz catheter, pressure-volume data were obtained from the right ventricle. Ten patients undergoing coronary revascularization served as controls. RESULTS: Twenty hearts were transplanted, seven of them as heart and lung blocks. Right ventricular end-systolic elastance (E(es)) was lower in BD donors compared with controls (mean 0.28 vs. 0.46 mm Hg/mL, P< or =0.01). There was no difference in terms of Swan-Ganz derived data between the BD subgroups, but E(es) was lower in groups 2 and 3 (P=0.04). Eight patients died within a year (four from graft failure), and they had a donor heart E(es) significantly lower than that of survivors (mean 0.20 vs. 0.33 mm Hg/mL, P=0.01). CONCLUSION: Hearts from BD donors have subclinical right ventricular impairment in contractility. E(es), a load-independent measure of contractile function, seems to be inversely correlated with empirical use of noradrenaline in the donor and with recipient survival at 1 year. This has implications for refining donor selection and management.

Adult↗
PubMed1992

Chromosomal targeting of replicating plasmids in the yeast Hansenula polymorpha.

Using an optimized transformation protocol we have studied the possible interactions between transforming plasmid DNA and the Hansenula polymorpha genome. Plasmids consisting only of a pBR322 replicon, an antibiotic resistance marker for Escherichia coli and the Saccharomyces cerevisiae LEU2 gene were shown to replicate autonomously in the yeast at an approximate copy number of 6 (copies per genome equivalent). This autonomous behaviour is probably due to an H. polymorpha replicon-like sequence present on the S. cerevisiae LEU2 gene fragment. Plasmids replicated as multimers consisting of monomers connected in a head-to-tail configuration. Two out of nine transformants analysed appeared to contain plasmid multimers in which one of the monomers contained a deletion. Plasmids containing internal or flanking regions of the genomic alcohol oxidase gene were shown to integrate by homologous single or double cross-over recombination. Both single- and multi-copy (two or three) tandem integrations were observed. Targeted integration occurred in 1-22% of the cases and was only observed with plasmids linearized within the genomic sequences, indicating that homologous linear ends are recombinogenic in H. polymorpha. In the cases in which no targeted integration occurred, double-strand breaks were efficiently repaired in a homology-independent way. Repair of double-strand breaks was precise in 50-68% of the cases. Linearization within homologous as well as nonhomologous plasmid regions stimulated transformation frequencies up to 15-fold.

DNA Repair↗
PubMed2025

Systematic Optimization Enables Near-Perfect In Vitro Transformation Efficiencies for Spirodela polyrhiza (Greater Duckweed).

The in vitro transformation of plants, or the delivery of foreign genetic material that is incorporated into their genomes, represents a powerful tool both for elucidating genotype-phenotype relationships and for generating plant cultivars which have desirable traits for agriculture and/or biotechnological applications. However, outside of a few model species, the processes involved in transformation are often inefficient and can take months to perform for many plant species, with several bottlenecks occurring at the different stages of calli induction, genetic transfection, and plant regeneration. While duckweeds - aquatic monocots whose species include some of the smallest and fastest-growing flowering plants on the planet - have distinguished themselves with several emerging biotechnological applications, they too are the subject of conflicting reports regarding their transformation potential and ability to be genetically manipulated. Here, we synthesized and optimized the protocols for in vitro transformation of duckweed Spirodela polyrhiza (Greater Duckweed) from start-to-finish: achieving >90% - 100% efficiencies for each of calli induction; transient and stable genetic transformation; visual marker-free selection of transformants; and regeneration of genetically modified plants with stable transgene expression for over 100 generations - and which in S. polyrhiza can be achieved over the course of weeks instead of months. The integrated, streamlined approaches for all stages of in vitro transformation overcome many bottlenecks and can help to pave the way for high-throughput functional genomics studies and synthetic biology applications in this biotechnologically-important species.

CRISPR/Cas9↗
PubMed1994

Computing genetic similarity coefficients from RAPD data: correcting for the effects of PCR artifacts caused by variation in experimental conditions.

The production of informative random amplified polymorphic DNA (RAPD) markers using PCR and a single primer is often accompanied by the generation of artifactual (noninformative) bands as well. When RAPD data are used to compute genetic similarity coefficients, these artifacts (false positives, false negatives, or both) can cause large biases in the numerical values of the coefficients. As a result, some workers have been reluctant to use RAPD markers in the estimation of genetic similarities. Artifactual bands are of two types: those caused by variation in experimental conditions, and those caused by characteristics of the DNA to be amplified. A procedure is described that allows for correction of the bias caused by the first type of artifact, providing that replicate DNA samples have been extracted, amplified, and scored. The resulting data are used to obtain an estimate of the proportion of false-positive and false-negatives bands. These values are then used to correct the bias in the computed similarity coefficients. Two examples are given, one in which bias correction is critical to the results, and one in which it is less important. The maximum percent bias, computed from the estimated proportions of false positives and false negatives in the RAPD data set, is proposed as a criterion for determining whether bias correction of the similarity coefficients is required or not. Although all reasonable efforts should be made to optimize PCR protocols to eliminate artifactual bands, when this is not possible, the methods described allow RAPD markers to compute genetic similarities reliably and accurately, even when artifactual bands resulting from variation in experimental conditions are present.

Artifacts↗
PubMed2001

Low-entanglement remote state preparation.

An outer bound on the low-entanglement remote state preparation ebits vs bits tradeoff curve [Bennett et al., quant-ph/0006044, 2000] is found using techniques of classical information theory. We show this bound to be optimal among an important class of protocols and conjecture optimality even without this restriction.

Journal Article↗
PubMed2004

Feedback control in a collective flashing ratchet.

An ensemble of Brownian particles in a feedback controlled flashing ratchet is studied. The ratchet potential is switched on and off depending on the position of the particles, with the aim of maximizing the current. We study in detail a protocol which maximizes the instant velocity of the center of mass of the ensemble at any time. This protocol is optimal for one particle and performs better than any periodic flashing for ensembles of moderate size, but is defeated by a random or periodic switching for large ensembles.

Journal Article↗