Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “protocol optimization”

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 1,261 records · Page 70Linked to original sources
PubMed2005

A monoclonal antibody with potential for aiding non-invasive prenatal diagnosis: utility in screening of pregnant women at risk of preeclampsia.

The development of a non-invasive method of prenatal diagnosis in maternal blood has been the goal of our investigations during the last years. We have developed several anti-CD71 monoclonal antibodies and optimized a protocol for the isolation of nucleated red blood cells (NRBC) from peripheral maternal blood. The enhanced traffic of fetal erythroblasts into the maternal circulation in preeclampsia has been investigated by several groups. In this study, we compared one of our antibodies, 2F6.3, with a commercial anti-CD71 antibody in blood samples from pregnant women suffering pregnancy-induced hypertension (PIH) and in a control group of pregnant women without clinical features suggestive of an increased risk of developing preeclampsia. The mAb 2F6.3, developed by our group, has succeeded in isolating a significantly higher number of erythroblasts with less maternal cell contamination than the commercial antibody in both women with PIH and in the control group (p<0.01; Wilcoxon Signed Ranks Test). Fluorescence in situ hybridization analysis also demonstrated that 2F6.3 is a better antibody for the isolation of fetal NRBC in maternal blood than the commercial anti-CD71 antibody.

Antibodies, Monoclonal↗
PubMed2005

Optimized real-time quantitative PCR measurement of male fetal DNA in maternal plasma.

BACKGROUND: Circulating fetal DNA (cfDNA) in maternal plasma has been measured to investigate its possible relationship with pregnancy-related disorders, including fetal trisomy 21 and preeclampsia. The circulating concentrations of single-copy fetal genes, however, are close to the detection limits of PCR methods. METHODS: We optimized a protocol for the real-time quantitative PCR amplification of the multicopy sequence DYS14 on the Y-chromosome. This was compared with an established real-time PCR assay for the single-copy SRY gene. RESULTS: By probit regression analysis, the measurements of male DNA by the DYS14 assay had a 10-fold lower detection limit (0.4 genome equivalents) than did measurements of SRY. For plasma samples from women in the first trimester of pregnancy, imprecision (CV) was 2%-22% when amplifying DYS14 compared with 26%-140% for SRY. CONCLUSIONS: The low copy numbers of fetal DNA in plasma of women in the first trimester of pregnancy cannot be measured precisely when targeting single-copy sequences. Better results are obtained by amplifying a sequence that is present in multiple copies per male genome.

Chromosomes, Human, Y↗
PubMed2005

Current issues in mechanical ventilation for respiratory failure.

The morbidity and mortality associated with respiratory failure is, to a certain extent, iatrogenic. Mechanical ventilation, although the mainstay of treatment for respiratory distress syndrome, can result in physical trauma to lung tissue (ventilator-induced lung injury [VILI]). Strategies to alleviate VILI are often termed lung-protective strategies and are aimed at reducing overstretching and shear stresses associated with repetitive alveolar collapse and reopening. Lower tidal volumes during ventilation, maintenance of positive-end expiratory pressure, and high-frequency ventilation are the best-studied lung-protective strategies that appear to reduce VILI. Faster withdrawal from mechanical ventilation could also improve outcomes and lower the costs associated with care. To enhance the success of weaning from mechanical ventilation, the cooperative efforts of physicians and respiratory therapists are needed. These efforts involve the initiation of spontaneous-breathing trials, implementation of systematic weaning protocols, and optimization of individual patient interventions.

Animals↗
PubMed2005

Biomedical informatics methods in pharmacogenomics.

Pharmacogenomics is the study of the genetic basis of individual variation in response to therapeutic agents. Pharmacogenomics may potentially affect on every step of health care and every drug treatment protocol. The optimal approach to pharmacogenomics in hypertension requires the integration of different disciplines, in which biomedical informatics plays an essential role. This chapter describes biomedical informatics methods used in dealing with key issues in pharmacogenomics. These key issues include the association between structure and function, the interaction between gene and drug, and the correlation between genotype and phenotype. Heterogeneous resources, including web sites, databases, and software analysis tools, are selected, organized, and integrated in practical methods to support these studies. Bioinformatics methods described in this chapter include genetic sequence searching, comparison, structural modeling, functional analysis, and systems biology studies, with emphasis on single-nucleotide polymorphism (SNP) analysis. Medical informatics methods such as disease and drug information and clinical terminology are also embraced in this chapter. This combination of both biological and medical informatics provides comprehensive methodologies to resolve complex problems in pharmacogenomics.

Computational Biology↗
PubMed2002

Cancer immunotherapy with interleukin-2-activated natural killer cells.

Natural killer (NK) cells are a subset of lymphocytes with a distinct morphologic appearance (large granular lymphocytes [LGL]) and the ability to kill virally infected and tumor targets but to spare most normal cells. NK cells respond to a variety of biologic agents, such as interleukin-2 (IL-2), or interferons, by upregulation of cytolytic, secretory, and proliferative functions. In cancer-bearing hosts, NK cells have been considered to be the major component of antitumor immunity responsible for rapid elimination of malignant cells from the blood. More recently, however, studies have demonstrated the ability of adoptively transferred, IL-2-activated NK cells to selectively localize into solid tumors tissue and to eliminate established tumors. While these findings indicate a role for NK cells in cancer immunotherapy, additional studies are needed in both animal models and in humans to optimize clinical protocols of cancer therapy based on these cells.

Adoptive Transfer↗
PubMed2001

Interaction between photodynamic therapy and BCG immunotherapy responsible for the reduced recurrence of treated mouse tumors.

Subcutaneous mouse EMT6 tumors were treated by individual or combined regimens of a single Bacillus Calmette-Guérin (BCG) vaccine administration and photodynamic therapy (PDT). Six clinically relevant photosensitizers characterized by different action mechanisms were used: Photofrin, benzoporphyrin derivative, tetra(m-hydroxyphenyl)chlorin (foscan), mono-L-aspartylchlorin e6, lutetium texaphyrin or zinc phthalocyanine. Irrespective of the type of photosensitizer used, the optimized BCG protocols improved the cure rate of PDT-treated tumors. This indicates that the interaction does not take place during the early phase of tumor ablation but at later events involved in preventing tumor recurrence. Beneficial effects on tumor cure were observed even when the BCG injection was delayed to 7 days after PDT. The accumulation of activated myeloid cells that markedly increases in tumors treated by Photofrin-based PDT was not additionally affected by BCG treatment. However, the incidence of immune memory T cells in tumor-draining lymph nodes that almost doubled at 6 days after Photofrin-PDT further increased close to three-fold with adjuvant BCG. This suggests that BCG immunotherapy amplifies the T-lymphocyte-mediated immune response against PDT-treated tumors. Since both these modalities are established for the treatment of superficial bladder carcinomas, use of their combination for this condition should be clinically tested.

Adjuvants, Immunologic↗
PubMed2006

Genetically modified dendritic cells for therapeutic immunity.

Dendritic cells are professional antigen presenting cells, which show an extraordinary capacity to initiate primary immune responses by stimulating T cells. This established function of dendritic cells has attracted much attention in efforts to develop useful vaccines for the treatment of cancer and infectious diseases. Designing effective strategies to generate clinical dendritic cell-based vaccine protocols remains a challenging field of research. The successful realization of immunotherapy utilizing dendritic cells will depend on modifications of these protocols to optimize the natural stimulatory properties of dendritic cells, such as genetic modification of dendritic cells. This review focuses on dendritic cell gene modifications for enhancing the multiple effector functions of dendritic cells, including viral and non-viral gene transfer into dendritic cells, and a variety of transferred genes, such as those encoding antigens, co-stimulatory molecules, cytokines, and chemokines.

Antigens↗
PubMedDate not supplied

Hypofractionated radiation therapy of brain masses in dogs: a retrospective analysis of survival of 83 cases (1991-1996).

There are few reports of radiation treatment for brain tumors in dogs, and the optimal treatment protocol has yet to be established. We completed a retrospective analysis of the survival times of a series of 83 dogs with intracranial masses that were treated by hypofractionated megavoltage radiation therapy. A total tumor dose of 38 Gray was given over 5 weeks as once weekly fractions via 3 perpendicular portals. The median survival time from the start of radiotherapy for the whole cohort was 43.7 weeks (range, 0.1-172 weeks). Extra-axial masses had a better survival time (49.7 weeks) than did other intracranial masses (intra-axial, 40.4 weeks; pituitary, 21.0 weeks). Delayed radiation toxicity was suspected as the cause of death or reason for euthanasia in 12 dogs. The hypofractionated radiation protocol resulted in survival times similar to those obtained using more conventional multifractionated regimens, and this protocol may be a useful, less intensive alternative treatment for brain tumors in dogs.

Adenoma↗
PubMed1999

Synthesis and automated detection of fluorescently labeled primer extension products.

Common methods for determination of transcription initiation sites are based on the extension of radioactively labeled primers or internal labeling of extension products with reverse transcriptase. Here, we describe a modification of this procedure, adapted for automated detection of extension products by using fluorescent primers and an optimized reaction protocol. This facilitates high-throughput screening to detect mRNA start-points of large DNA regions and significantly reduces the time required to determine sites of transcription initiation.

Bacteriophages↗
PubMed1999

Nonradioactive labeling of large DNA fragments for genome walking, RFLP and northern blot analysis.

In this report, we present a simple nonradioactive labeling procedure for DNA fragments of high specific labeling density that can be used for a variety of applications. The protocol is based on the universal mono-functional platinum reagent for chemical digoxigenin (DIG) labeling of nucleic acids. The labeling protocol was optimized for large DNA templates as complete bacterial artificial chromosomes (BAC). Variations of incubation time and temperature improved the labeling density such that about 30% of the nucleotides were DIG-modified within 30 min. Furthermore, the refined procedure generates in a single-tube reaction and without prior digestion-labeled DNA fragments of 0.5-4.0 kb from a 130-kb template. Hybridization experiments were performed on Southern and northern blots and allowed the detection of single copy genes in 2.5 micrograms genomic DNA from Arabidopsis thaliana, which has a haploid genome size of 0.13 pg (ca. 120 Mb) and medium expressed transcripts from 0.8 microgram poly(A)+ RNA, respectively. The extremely high specific labeling density, the stability and the universal application of the probe generated with the platinum reagent makes this method a useful alternative to classical radioactive nuclei acids labeling techniques.

Alleles↗
PubMed2004

Imaging drug resistance with radiolabeled molecules.

A major obstacle to successful cancer chemotherapy is drug resistance. Multidrug resistance (MDR) is often seen with chemotherapeutic agents such as anthracycline derivatives, vinca alkaloids and taxanes. Multiple aspects of cellular biochemistry have been implicated in the MDR process. Cellular mechanisms of resistance are due to the presence of efflux pumps, P-glycoprotein (P-gp) and multiple resistance-associated protein (MRP), which belong to the ATP-binding cassette (ABC) family of transporters. Another form of drug resistance is involved in the chemotherapy of cancers with alkylating agents such as nitrosourea derivatives and nitrogen mustards. The cytotoxicity of these agents is primarily due to alkylation of the DNA guanine residues at their O6-position, which leads, via a cascade of events, to DNA strand breaks. The DNA repair protein, alkylguanine-DNA alkyl transferase (AGT) removes the alkyl groups from the lesions stoichiometrically to a cysteine in its active site. This process is irreversible and results in the degradation of the protein and its recovery is entirely from de novo synthesis. Noninvasive methodologies for monitoring the transport activity of these efflux pumps and determining tumor content of AGT could serve as critical tools for optimizing chemotherapeutic protocols on a patient-specific basis and gaining an understanding of the dynamics of resistance in living patients. In this review, we will describe the efforts made to date to synthesize radioactive probes of chemotherapy resistance and their use to quantitate these transporters and DNA repair protein by radionuclide imaging.

ATP-Binding Cassette Transporters↗
PubMed2004

HIV-specific CD8+ T cells: serial killers condemned to die?

An increasing body of evidence supports a key role for cytotoxic CD8+ T cells (CTL) in controlling HIV infection. Although a vigorous HIV-specific CD8+ T cell response is raised during the primary infection, these cells ultimately fail to control virus and prevent disease progression. The failure of CTL to control HIV infection has been attributed to a number of strategies HIV employs to evade the immune system. Recently, intrinsic defects in the CTL themselves have been proposed to contribute to the failure of CTL to control HIV. HIV-specific CD8+ T cells differ in their effector/memory phenotype from other virus-specific CD8+ T cells indicating that their differentiation status differs. This altered differentiation may affect effector functions as well as homing properties of these cells. Other studies have indicated that activation of HIV-specific CTL may be impaired and this contributes to their dysfunction. The effector function of these CTL may also be affected. There are conflicting reports about their ability to kill, whereas IFNgamma production does not appear to be impaired in these cells. In this review we focus on recent work indicating that apoptosis may be an important mechanism through which HIV evades the CTL response. In particular, HIV-specific CD8+ T cells are highly susceptible to CD95/Fas-induced apoptosis. This leads to the hypothesis that virus-specific cytotoxic T cells can be eliminated upon binding CD95L/FasL on HIV-infected cells. Understanding the intrinsic defects of CTL in HIV infection could lead to new therapeutic strategies and optimized vaccination protocols that enhance the HIV-specific cytotoxic response.

Apoptosis↗
PubMed1993

Interstitial MR lymphography with iron oxide particles: results in tumor-free and VX2 tumor-bearing rabbits.

OBJECTIVE: MR lymphography with interstitial injection of superparamagnetic iron oxide particles was optimized in normal rabbits by investigating the pattern of signal reduction in lymph nodes as a function of dose and time after administration. The optimized examination procedure was then used in a rabbit tumor model to study the potential of superparamagnetic iron oxide--enhanced MR lymphography in the detection of metastatic lymph node involvement. MATERIALS AND METHODS: The popliteal and iliac lymph nodes of 18 normal rabbits were imaged to study the dose response and time course of the effect of the contrast agent. For the dose response study, six doses of 2-50 mumol of iron per extremity were administered to three animals per dose, and MR images were obtained before and 12 hr after administration. For the time course study, 20 mumol of iron per extremity was administered to four animals, and images were obtained before and 6 hr to 42 days after administration. VX2 tumor-bearing rabbits were examined 12 hr after administration of 20 mumol (10 animals) and 50 mumol (three animals) of iron per extremity. Superparamagnetic iron oxide was injected into the foot pad of the hind limb. T1-, T2-, and proton density-weighted MR images were obtained with a 1.5-T unit. RESULTS: In normal rabbits, a profound and homogeneous loss of signal intensity was found with doses of 2-5 mumol of iron per extremity in popliteal lymph nodes and with doses of 20-30 mumol in the iliac lymph nodes. Superparamagnetic iron oxide caused maximal loss of signal intensity in both popliteal and iliac lymph nodes 12 hr after administration. In tumor-bearing rabbits, different degrees of metastatic displacement of lymph nodes were discernible, and even small metastases (3 mm in diameter) could be visualized when using the optimized examination protocol and the proton density-weighted spin-echo sequence. CONCLUSION: We conclude that interstitial MR lymphography with superparamagnetic iron oxide enables the detection of lymph node metastases and therefore is a promising technique for improved diagnostic imaging of lymph nodes in the staging of tumors.

Animals↗
PubMed2003

Four-channel multidetector CT in facial fractures: do we need 2 x 0.5 mm collimation?

OBJECTIVE: Our aim was to optimize acquisition protocols and multiplanar reformation algorithms for the evaluation of facial fractures using multidetector CT (MDCT) and to determine whether 2 x 0.5 mm collimation is necessary. MATERIALS AND METHODS: A cadaveric head with artificial blunt facial trauma was examined using a four-channel MDCT scanner. The influence of acquisition parameters (collimation, 2 x 0.5 mm, 4 x 1 mm, 4 x 2.5 mm; tube current, 120 mAs, 90 mAs, 60 mAs), image reconstruction algorithms (standard vs ultra-high-resolution modes; reconstructed slice thicknesses, 0.5 mm, 1 mm, 3 mm; increment, 0.3 mm, 0.6 mm, 1.5 mm), and reformation algorithms (slice thicknesses, 0.5 mm, 1 mm, 3 mm; overlap, 0.5 mm, 1 mm, 3 mm) on detectability of facial fractures in multiplanar reformations with MDCT was analyzed. RESULTS: Fracture detection was significantly higher with thin multiplanar reformations (0.5 and 0.5 mm, 1 and 0.5 mm, and 1 and 1 mm) (p < or = 0.014) acquired with 2 x 0.5 mm collimation (p < or = 0.046) in ultra-high-resolution mode (p < 0.0005) with 120 mAs (p < or = 0.025). Interobserver variability showed very good agreement (kappa > or = 0.942). Non-ultra-high-resolution mode, lower milliampere-seconds, and thick multiplanar reformations (3 and 0.5 mm, 3 and 1 mm, and 3 and 0.5 mm) showed significantly decreased fracture detectability. CONCLUSION: Although thin multiplanar reformations obtained from thin collimation (2 x 0.5 mm) are statistically superior for the detection of subtle fractures, 4 x 1 mm collimation is sufficient for routine diagnostic evaluation. Ultra-high-resolution mode with 120 mAs is mandatory for detection of clinically relevant fractures.

Aged↗
PubMed2001

Vaccine against Helicobacter pylori?

The burden of illness caused by Helicobacter pylori is considerable, and there is little doubt that a therapeutic vaccine would be very useful. However, the merits of using a prophylactic vaccine to eradicate H. pylori from the human population are debatable in light of arguments that it may act as a beneficial commensal organism for many of its human hosts. With regard to the practical side of designing vaccines against H. pylori, animal models have shown us that both therapeutic and prophylactic vaccines are feasible despite the fact that, as in human hosts, immune responses raised naturally to H. pylori cannot eradicate the infection. Many advances have been made in developing optimal vaccination protocols, and recently the first human trial of a therapeutic vaccine, which showed limited but nevertheless encouraging results, was published. However, a possibly quite serious problem is that prophylactic and perhaps also therapeutic vaccines may themselves cause disease.

Animals↗
PubMed2006

Autologous adult human Schwann cells genetically modified to provide alternative cellular transplants in peripheral nerve regeneration.

OBJECT: The purpose of this study was to provide genetically modified adult human Schwann cells as tools for cell transplantation in peripheral nerve repair. The application of transfected autologous Schwann cells overexpressing regeneration-promoting proteins, for example, neurotrophic or growth factors, is a promising approach in the aforementioned context. To achieve an optimal clinical outcome, it is highly important to perform enrichment, genetic modification, and retransplantation of cells in a short time. METHODS: To enable the development of these autologous cellular prostheses, the authors tested the properties of adult human Schwann cells obtained from differently treated human peripheral nerve biopsy samples. The use of "cold jet," a fast and effective enrichment procedure, as well as selective, serum-free culture conditions, resulted in very pure adult human Schwann cell cultures. Using an optimized electroporation protocol, as many as 48.4% of adult human Schwann cells were successfully transfected. CONCLUSIONS: The authors present a very fast protocol to establish adult human Schwann cell cultures that demonstrably express plasmid proteins after plasmid DNA insertion by electroporation. These autologous human Schwann cells transfected to enhance the endogenous production of regeneration-supporting proteins will likely constitute a major component of tissue-engineered peripheral nerve grafts.

Adolescent↗
PubMed2004

Provision, uptake and cost of cardiac rehabilitation programmes: improving services to under-represented groups.

OBJECTIVES: To estimate UK need for outpatient cardiac rehabilitation, current provision and identification of patient groups not receiving services. To conduct a systematic review of literature on methods to improve uptake and adherence to cardiac rehabilitation. To estimate cost implications of increasing uptake of cardiac rehabilitation. DATA SOURCES: Hospital Episode Statistics (England). Hospital Inpatient Systems (Northern Ireland). Patients Episode Database for Wales. British Association for Cardiac Rehabilitation/British Heart Foundation surveys. Cardiac rehabilitation centres. Patients from general hospitals. Electronic databases. REVIEW METHODS: The study analysed hospital discharge statistics to ascertain the population need for outpatient cardiac rehabilitation in the UK. Surveys of cardiac rehabilitation programmes were conducted to determine UK provision, uptake and audit activity, and to identify local interventions to improve uptake. Data were also examined from a trial estimating eligibility for cardiac rehabilitation and non-attendance. A systematic review of interventions to improve patient uptake, adherence and professional compliance in cardiac rehabilitation was conducted. Estimated costs of improving uptake were identified from national survey, systematic review and sampled cardiac rehabilitation programmes. RESULTS: In England, Wales and Northern Ireland nearly 146,000 patients discharged from hospital with primary diagnosis of acute myocardial infarction, unstable angina or following revascularisation were potentially eligible for cardiac rehabilitation. In England in 2000, 45-67% of these patients were referred, with 27-41% attending outpatient cardiac rehabilitation. If all discharge diagnoses of ischaemic heart disease were considered, nearly 299,000 patients would be potentially eligible and in England rates of attendance and referral would be 22-33% and 13-20% respectively. Rates of referral and attendance were similar in Wales, but somewhat lower in Northern Ireland. It was found that referral and attendance of older people and women at cardiac rehabilitation tended to be low. It was also suggested that patients from ethnic minorities and those with angina or heart failure were less likely to be referred to or join programmes. A wide range of local interventions suggested awareness of the problem of uptake. In an NHS-funded randomised controlled trial, possibly representing more optimal protocol-led care, medical and nursing staff identified 73-81% of patients with acute myocardial infarction as eligible for cardiac rehabilitation. Excluded patients tended to be older with more severe presentation of cardiac disease. Experiences of patients suggested that uptake may be improved by addressing issues of motivation and relevance of rehabilitation to future well-being, co-morbidities, site and time of programme, transport and care for dependents. Systematic review of studies supported the use of letters, pamphlets or home visits to motivate patients and the use of trained lay visitors. Self-management techniques showed some value in promoting adherence to lifestyle changes. Studies examining professional compliance found that professional support for practice nurses may have value in the coordination of postdischarge care. Average costs in 2001 of cardiac rehabilitation to the health service per patient completing a cardiac rehabilitation programme were about GBP350 (staff only) and GBP490 (total). If services were modelled on an intermediate multidisciplinary configuration with three to five key staff, approximately 13% more patients could be treated with the same budget. Depending on staffing configuration an approximate 200-790% budget increase would be required to provide cardiac rehabilitation to all potentially eligible patients. CONCLUSIONS: Provision of outpatient cardiac rehabilitation in the UK is low and little is known about the capacity of cardiac rehabilitation centres to increase this provision. There is an uncoordinated approach to audit data collection and few interventions aimed at improving the situation have been formally evaluated. Motivational communications and trained lay volunteers may improve uptake of cardiac rehabilitation, as may self-management techniques. Experience of low-cost interventions and good practice exists within rehabilitation centres, although cost information frequently is not reported. Increased provision of outpatient cardiac rehabilitation will require extra resources. Further trials are required to compare the cost-effectiveness of comprehensive multidisciplinary rehabilitation with simpler outpatient programmes, also research is needed into economic and patient preference studies of the effects of different methods of using increased funding for cardiac rehabilitation. An evaluation of a range of interventions to promote attendance in all patients and under-represented groups would also be useful. The development of standards is suggested for audit methods and for eligibility criteria, as well as regular and comprehensive data collection to estimate the need for and provision of cardiac rehabilitation. Further areas for intervention could be identified through qualitative studies, and the extension of low-cost interventions and good practice within rehabilitation centres. Regularly updated systematic reviews of relevant literature would also be useful.

Cost-Benefit Analysis↗
PubMed2001

Comparison of TaqMan Salmonella amplification/detection kit with standard culture procedure for detection of Salmonella in meat samples.

We evaluated the TaqMan PCR Salmonella amplification/detection kit (PE Applied Biosystems) for rapid detection of Salmonella from a variety of meat samples. This system uses the 5' nuclease activity of Taq DNA polymerase, which digests an internal fluorogenic probe to monitor the amplification of the target gene. The detection sensitivity of the kit, using 2 kinds of DNA extraction protocols, was compared with that obtained with 4 protocols of official culture methods. A total of 98 meat samples (16 raw beef, 31 pork and 51 chicken) were tested. The results of the TaqMan PCR method and the combined results of the 4 cultural protocols showed excellent agreement. However, no single culture protocol showed optimal recovery of Salmonella comparable to the PCR method. These results suggest that the TaqMan PCR method is a reliable and rapid method useful for detecting Salmonella in meat products.

Food Microbiology↗