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Biomedical subjects

R K Martin

Publications and source records attributed to R K Martin.

At least 19 recordsLinked to original sources

Who should manage continuous renal replacement in the intensive care setting? A nursing viewpoint.

Since its inception, continuous renal replacement therapy (CRRT) has been performed in critical care units with or without the involvement of nephrology nursing support (1,2). It is apparent that the issue of providing care to patients requiring this therapy is not so much a debate on the nursing control of CRRT, but a focused discussion on the nursing management and delivery of care to the patient receiving CRRT in the intensive care setting. Although the choice of a nursing care model for CRRT is dependent on many clinical and organisational factors, the use of one nursing specialty to deliver CRRT care can leave gaps in practice. The Joint or Collaborative Nephrology/critical care nursing model brings the highest level of nursing expertise to the bedside. The joint model tends to promote collaboration between two distinct nursing specialties, with opportunities for setting joint standards and promoting research. With this in mind, this discussion will examine some of the factors affecting structuring of nursing care, describe nursing models currently in use, compare the attributes of each, and conclude which model is preferred for the delivery of nursing care for CRRT.

Clinical Competence↗

Serial analysis of gene expression (SAGE) in Plasmodium falciparum: application of the technique to A-T rich genomes.

The advent of high-throughput methods for the analysis of global gene expression, together with the Malaria Genome Project open up new opportunities for furthering our understanding of the fundamental biology and virulence of the malaria parasite. Serial analysis of gene expression (SAGE) is particularly well suited for malarial systems, as the genomes of Plasmodium species remain to be fully annotated. By simultaneously and quantitatively analyzing mRNA transcript profiles from a given cell population, SAGE allows for the discovery of new genes. In this study, one reports the successful application of SAGE in Plasmodium falciparum, 3D7 strain parasites, from which a preliminary library of 6880 tags corresponding to 4146 different genes was generated. It was demonstrated that P. falciparum is amenable to this technique, despite the remarkably high A-T content of its genome. SAGE tags as short as 10 nucleotides were sufficient to uniquely identify parasite transcripts from both nuclear and mitochondrial genomes. Moreover, the skewed A-T content of parasite sequence did not preclude the use of enzymes that are crucial for generating representative SAGE libraries. Finally, a few modifications to DNA extraction and cloning steps of the SAGE protocol proved useful for circumventing specific problems presented by A-T rich genomes.

Animals↗

Co-ordinated programme of gene expression during asexual intraerythrocytic development of the human malaria parasite Plasmodium falciparum revealed by microarray analysis.

Plasmodium falciparum is a protozoan parasite responsible for the most severe forms of human malaria. All the clinical symptoms and pathological changes seen during human infection are caused by the asexual blood stages of Plasmodium. Within host red blood cells, the parasite undergoes enormous developmental changes during its maturation. In order to analyse the expression of genes during intraerythrocytic development, DNA microarrays were constructed and probed with stage-specific cDNA. Developmental upregulation of specific mRNAs was found to cluster into functional groups and revealed a co-ordinated programme of gene expression. Those involved in protein synthesis (ribosomal proteins, translation factors) peaked early in development, followed by those involved in metabolism, most dramatically glycolysis genes. Adhesion/invasion genes were turned on later in the maturation process. At the end of intraerythrocytic development (late schizogony), there was a general shut-off of gene expression, although a small set of genes, including a number of protein kinases, were turned on at this stage. Nearly all genes showed some regulation over the course of development. A handful of genes remained constant and should be useful for normalizing mRNA levels between stages. These data will facilitate functional analysis of the P. falciparum genome and will help to identify genes with a critical role in parasite progression and multiplication in the human host.

Animals↗

Technical assessment of the affymetrix yeast expression GeneChip YE6100 platform in a heterologous model of genes that confer resistance to antimalarial drugs in yeast.

The advent of high-density gene array technology has revolutionized approaches to drug design, development, and characterization. At the laboratory level, the efficient, consistent, and dependable exploitation of this complex technology requires the stringent standardization of protocols and data analysis platforms. The Affymetrix YE6100 expression GeneChip platform was evaluated for its performance in the analysis of both global (6,000 yeast genes) and targeted (three pleiotropic multidrug resistance genes of the ATP binding cassette transporter family) gene expression in a heterologous yeast model system in the presence and absence of the antimalarial drug chloroquine. Critical to the generation of consistent data from this platform are issues involving the preparation of the specimen, use of appropriate controls, accurate assessment of experiment variance, strict adherence to optimized enzymatic and hybridization protocols, and use of sophisticated bioinformatics tools for data analysis.

ATP-Binding Cassette Transporters↗

The role of the transplant advanced practice nurse: a professional and personal evolution.

Rapid changes in health care toward cost-effective managed care have resulted in the evolution of the transplant advanced practice nurse to address the care needs of a specific patient population. This article describes the evolution of this role at one institution. The specific responsibilities of the transplant advanced practice nurse are delineated, and phases of development for the experienced clinical nurse specialist entering the role as a novice nurse practitioner are identified and discussed.

Cost-Benefit Analysis↗

Organ transplantation: the role of the acute care nurse practitioner across the spectrum of health care.

Organ transplantation has become an accepted treatment method for a multitude of conditions once deemed untreatable. Increasing numbers of patients are treated by nurse practitioners before and after transplantation in a variety of settings, such as primary and specialty care clinics, urgent care, critical care units, and long-term care facilities. This article outlines the role of the acute care nurse practitioner in the management of adult transplant recipients in a variety of health care settings and reviews the problems and interventions specific to this patient group. Emphasis is placed on the referral process and collaboration with the transplantation team in patient management.

Acute Disease↗

Preservation of nucleic acids for polymerase chain reaction after prolonged storage at room temperature.

A guanidinium isothiocyanate (GITC) lysis solution was evaluated for its efficacy in preserving nucleic acids for subsequent analysis after prolonged storage at room temperature. Aliquots of thyroid crude cell lysates were stored at 22 degrees C for 8 weeks in the GITC solution. Crude cell lysates stored for 2 weeks in the GITC solution consistently provided adequate RNA and DNA for reverse transcriptase-polymerase chain reaction (RT-PCR) and PCR, respectively, of beta 2-microglobulin, thyroid stimulating hormone receptor (TSHR), and thyroglobulin. Interestingly, the thyroglobulin and TSHR messages from thyroid fine needle aspirate samples were detected by RT-PCR only when stored at room temperature for less than 1 week, whereas the RT-PCR product for beta 2-microglobulin was detectable in 95% of the samples at 3 months. In addition, thyroglobulin DNA was amplified by PCR in nearly all samples stored at 22 degrees C for 2 months. These data suggest that GITC solutions can be used to preserve nucleic acids in most circumstance until transported to laboratories equipped and staffed with appropriate resources.

Biopsy, Needle↗

The use of the polymerase chain reaction to detect bacteria in amniotic fluid in pregnancies complicated by preterm labor.

OBJECTIVES: The purpose of this investigation was to determine the feasibility of using the polymerase chain reaction to detect bacteria in amniotic fluid and to compare pregnancy outcomes in subsets of women categorized by amniotic fluid culture, polymerase chain reaction, and interleukin-6 findings. STUDY DESIGN: Amniotic fluid from 54 pregnancies with preterm labor and no clinical evidence of intraamniotic infection was evaluated with use of the polymerase chain reaction, interleukin-6, and bacterial culture. Gestational age, newborn weight, and time between amniocentesis and delivery were compared between subsets of women categorized by these tests. RESULTS: With use of the polymerase chain reaction <100 bacteria per milliliter could be detected in amniotic fluid. A total of 55.5% of the amniotic fluid samples were polymerase chain reaction positive, whereas 9.2% of culture results were positive. Birth weights and gestational age at delivery were less and time from amniocentesis to delivery was shorter in the polymerase chain reaction-positive group (p < 0.05). Nine samples (15%) had elevated interleukin-6 concentrations; of these, six were polymerase chain reaction positive. CONCLUSIONS: The polymerase chain reaction is a sensitive means of detecting bacteria in amniotic fluid. These results provide further evidence of an association between preterm delivery and intraamniotic infection. Not all amniotic fluid samples with elevated interleukin-6 levels have bacteria detectable by the polymerase chain reaction. We anticipate that the polymerase chain reaction will provide another avenue for the detection of bacteria in amniotic fluid.

Adult↗

Who should manage CRRT in the ICU? The nursing viewpoint.

Continuous renal replacement therapy (CRRT) is performed in critical care units around the world with various levels of involvement from critical care and nephrology nurses. In this article, factors affecting the delivery of nursing care and the particular expertise nephrology and critical care nurses have in the area of CRRT are examined. Based on this discussion, three proposed models of nursing care for the patient receiving CRRT are discussed: the Nephrology Nursing Model, the Critical Care Model, and the Collaborative Model. Based on related research findings and a comparison of the models, the Collaborative Model is the preferred one, as it brings the highest level of expertise directly to the patient. For the Collaborative Model to work, a framework for collaboration and a high degree of commitment from both specialties must be maintained.

Humans↗

Detection of ptc in archival formalin-fixed, paraffin-embedded tissues. Comparison of radiolabeled DNA hybridization and direct incorporation of digoxigenin-11-dUTP into RT-PCR products.

Archival pathological specimens are a source of RNA and DNA for clinical surveillance or retrospective studies. We employed a modification of the acid guanidium thiocyanate-phenol-chloroform extraction method for the recovery of total RNA from formalin-fixed, paraffin-embedded neoplastic thyroid tissue. The extracted RNA was used for reverse transcription of ptc and subsequent amplification of the complementary DNA (cDNA) by the reverse transcription-polymerase chain reaction (RT-PCR). In lieu of 32P-labeled DNA for hybridization studies, we supplemented the nucleotide pool in the amplification reaction with a modified pyrimidine, digoxigenin-11-dUTP. Digoxigenin-11-dUTP was incorporated directly into the PCR product, eliminating the need for hybridization, posthybridization washes, and prolonged autoradiography. These products were resolved by electrophoresis on agarose gels, Southern blotted to nylon membranes, and rapidly detected by chemiluminescence. This nonradioisotopic method has expedited and reduced the cost for molecular investigations with archival pathological specimens by providing equal sensitivity to or greater sensitivity than that of DNA-labeled radionuclides without the associated biological hazards.

Carcinoma, Papillary↗

Amplification of pfmdr 1 associated with mefloquine and halofantrine resistance in Plasmodium falciparum from Thailand.

Drug resistance in Plasmodium falciparum is an expanding problem in most endemic areas. Recent studies have suggested the potential involvement of genes in the MDR gene family in resistance to quinoline-containing compounds in P. falciparum. In this study a molecular analysis of pfmdr 1 in recent isolates from Thailand was done (1) to further examine the role of pfmdr 1 in drug-resistant isolates and (2) to examine the reported association of pfmdr 1 intragenic alleles and chloroquine resistance. Most of the isolates (10 of 11) were resistant to all compounds tested. Analysis of pfmdr 1 revealed an apparent association between increased gene copy number and increased level of expression of pfmdr 1 and decreased susceptibility to mefloquine and halofantrine. Sequence analysis of pfmdr 1 in these isolates revealed no association of intragenic alleles with chloroquine resistance.

Animals↗

Fluoxetine hydrochloride enhances in vitro susceptibility to chloroquine in resistant Plasmodium falciparum.

The emergence of chloroquine resistance in Plasmodium falciparum has necessitated the development of alternate strategies for chemotherapy and chemoprophylaxis. One approach has been the identification of drugs that do not possess any intrinsic antimalarial activity when used alone but that potentiate the effect of currently available antimalarial drugs, such as chloroquine. We identified fluoxetine hydrochloride (Prozac), a commonly prescribed antidepressant, as another resistance modulator for drug-resistant P. falciparum. Studies with chloroquine-resistant clones and isolates from various geographical locations confirmed our initial observations with a chloroquine-resistant P. falciparum clone, W2. Fluoxetine concentrations of 500 nM were found to effectively modulate chloroquine resistance by 66% in clone W2. In comparison, verapamil at similar concentrations was observed to modulate chloroquine resistance in clone W2 by 61%. Neither fluoxetine nor verapamil was observed to possess any innate antimalarial activity. These data augment the current description of the chloroquine resistance phenotype and may provide additional insights into lead-directed synthesis of new antimalarial drugs.

Animals↗

Structural diversity in the Plasmodium falciparum merozoite surface antigen 2.

Antigens associated with the surface of merozoites of the malaria parasite Plasmodium falciparum are directly accessible to immune attack and therefore are prime vaccine candidates. We have previously shown that one of the two known merozoite surface antigens (merozoite surface antigen 2; MSA-2) exhibits considerable sequence and antigenic diversity in different isolates. The sequences of MSA-2 from three isolates revealed a central domain composed of repeats that vary in number, length, and sequence, flanked in turn by nonrepetitive variable sequences and by conserved N- and C-terminal domains. We report here the sequences of a further four MSA-2 alleles, containing repetitive sequences that are related but not identical to each other. The seven alleles of MSA-2 can be divided into two distinct allele families on the basis of nonrepetitive sequences. Hybridization studies with repeat probes indicated that all of the 44 P. falciparum isolates examined contained repeat regions similar to those defined in known MSA-2 sequences.

Amino Acid Sequence↗

Characterization of a Leishmania isolate from the rodent host Neotoma micropus collected in Texas and comparison with human isolates.

We report the biological and biochemical parameters of Leishmania parasites (MNEO/US/90/WR972) isolated from a rodent host, Neotoma micropus, collected in Texas. Footpad inoculations of WR972 promastigotes into BALB/c mice and Syrian hamsters resulted in ulcerating lesions six and eight weeks post-inoculation, respectively. Using monoclonal antibody-stained touch preparations, amastigotes were found in the liver of both laboratory hosts. Infection of J774 macrophages with WR972 promastigotes supported the growth of amastigotes for 12 days at 35 degrees C. The WR972 parasite was identified by enzyme electrophoresis as L. mexicana. Isozyme comparison of WR972 with 42 L. mexicana isolates (from humans and rodents) from four different endemic areas, including Texas, suggest that these parasite populations are identical for approximately 97% of their genetic loci. Pulse field gel electrophoresis (PFGE) of WR972 resolved 18 chromosomes with a size range of 300- greater than 2,000 kb. The karyotype strongly resembles that of two other Texas L. mexicana isolates from humans. Taken together, the PFGE, hybridization, and isoenzyme data suggest that the wood rat isolate (WR972) is identical to parasites from human cutaneous lesions isolated in Texas and Central America. In addition, the biological characteristics of WR972, its infectivity of BALB/c mice and the Syrian hamster, and the potential of the isolate to infect, transform, and divide in J774 macrophages indicate that WR972 will be pathogenic in humans if transmission occurs. Health care providers should consider this possibility when studying the epidemiology and control of cutaneous leishmaniasis in Texas.

Animals↗

Characteristics of multidrug resistance in Plasmodium and Leishmania: detection of P-glycoprotein-like components.

Multidrug-resistance (MDR) in neoplastic cells is frequently characterized by the overexpression of P-glycoprotein (PGP), a 170 kDa transmembrane glycoprotein that binds multiple cytotoxic drugs as well as calcium channel antagonists. Chloroquine resistance in Plasmodium falciparum appears to be analogous to MDR in neoplastic cells, where the induction of resistance with one drug confers resistance to other structurally and functionally unrelated drugs. To test the hypothesis that chloroquine resistance in P. falciparum and antimony resistance in Leishmania is mediated by a similar mechanism of MDR in mammalian neoplastic cells, a PGP-specific monoclonal antibody (C219) was used to determine the presence of PGP genes in resistant and sensitive Plasmodium and Leishmania parasites by indirect immunofluorescence assays and Western blotting procedures. These PGP-like components were detected in both drug-sensitive and -resistant Plasmodium and Leishmania cells. A 40-42 kDa component was observed to be greater in a chloroquine-resistant P. berghei (C line) than in a chloroquine-susceptible P line. Differences observed between Pentostam-resistant and -sensitive Leishmania promastigote clones and isolates included the increased expression of 96-106 and 23-25 kDa peptides in drug-resistant L. enrietti, and increased amounts of two different peptides in two drug-resistant L. panamensis clones (i.e., 96-106 and 43-45 kDa in WR-746-CL4, and 53 and 23-25 kDa in kDa) in amastigotes as in MDR KB carcinoma cells (KB-V1). Comparative indirect immunofluorescent studies suggested that a correlation existed between the degree of antimony susceptibility and the concentration of the moiety recognized by C219 in two L. panamensis clones. Binding of the C219 monoclonal antibody to the PGP-like component of Leishmania was blocked by Pentostam, while the binding of C219 to multiple-drug resistant KB-V1 PGP was not inhibited by Pentostam, regardless of the PGP concentration. This suggests some degree of specificity in the binding of Pentostam to the Leishmania PGP-like components. In addition, these studies have demonstrated that drug-sensitive Leishmania accumulate two to five times more 125Sb-Pentostam than resistant clones.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Bundling--new products, new markets, low risk.

It has long been a marketing axiom that customers buy bundles of satisfaction, not products. It follows, then, that they'll respond to certain combinations of products and services--air conditioners with free installation, combinations of software packages, or season tickets with parking privileges. The difficulty is in devising the bundles that both appeal to consumers and give cost or demand enhancing benefits to the producer. Eppen, Hanson, and Martin argue that the best approach is to treat bundles not as marketing gimmicks but as new products. They offer seven guidelines for creating competitive bundles and a framework for implementing them.

Decision Making↗