Sequencing identifies a distinct signature of circulating microRNAs in early radiographic knee osteoarthritis.

OBJECTIVE: MicroRNAs act locally and systemically to impact osteoarthritis (OA) pathophysiology, but comprehensive profiling of the circulating miRNome in early vs late stages of OA has yet to be conducted. Sequencing has emerged as the preferred method for microRNA profiling since it offers high sensitivity and specificity. Our objective was to sequence the miRNome in plasma from 91 patients with early [Kellgren-Lawrence (KL) grade 0 or 1 (n = 41)] or late [KL grade 3 or 4 (n = 50)] symptomatic radiographic knee OA to identify unique microRNA signatures in each disease state. DESIGN: MicroRNA libraries were prepared using the QIAseq miRNA Library Kit and sequenced on the Illumina NextSeq 550. Counts were produced for microRNAs captured in miRBase and for novel microRNAs. Statistical, bioinformatics, and computational biology approaches were used to refine and interpret the final list of microRNAs. RESULTS: From 215 differentially expressed microRNAs (FDR < 0.01), 97 microRNAs showed an increase or decrease in expression in ≥85% of samples in the early OA group as compared to the median expression in the late OA group. Increasing this threshold to ≥95%, seven microRNAs were identified: hsa-miR-335-3p, hsa-miR-199a-5p, hsa-miR-671-3p, hsa-miR-1260b, hsa-miR-191-3p, hsa-miR-335-5p, and hsa-miR-543. Four novel microRNAs were present in ≥50% of early OA samples and had 27 predicted gene targets in common with the prioritized set of predicted gene targets from the 97 microRNAs, suggesting common underlying mechanisms. CONCLUSION: Sequencing of well-characterized patient cohorts produced unbiased profiling of the circulating miRNome and identified a unique panel of 11 microRNAs in early radiographic knee OA.

Early radiographic and functional outcomes of a cancellous titanium-coated tibial component for total knee arthroplasty.

BACKGROUND: Various surface coatings have been developed over the past decades to enhance fixation of cementless total knee arthroplasty (TKA). BIOFOAM(®) (MicroPort Orthopedics Inc., Arlington, TN, USA) is a novel cancellous titanium surface coating intended to increase both initial and long-term fixation. The purpose of this study was to investigate the early functional and radiographic outcomes of this coating used in a TKA application. MATERIALS AND METHODS: One hundred and four (104) primary TKAs in 85 subjects using BIOFOAM-coated tibial components were prospectively enrolled at four centers. Subjects were evaluated using Knee Society Scores and radiographic analysis at a minimum follow-up of 24 months. RESULTS: Knee Society Scores and flexion were all significantly improved at final follow-up compared to baseline. Radiographic analyses were satisfactory, with no progressive radiolucencies and only a single subject presenting with a radiolucency surrounding a tibial component. There were two revisions in the cohort: one for instability following a ruptured lateral collateral ligament and one for recurrent tibial insert dislocation. CONCLUSIONS: This is the first study to report clinical outcomes associated with the BIOFOAM coating used in a cementless TKA application. Early functional scores and radiographic analyses are promising, but further investigations are needed to confirm long-term clinical success with these components.

Adapting, piloting and evaluating complex public health interventions: lessons learned from the Nurse-Family Partnership in Canadian public health settings.

INTRODUCTION: The Nurse-Family Partnership (NFP) is a home-visit program for young and first-time, socially and economically disadvantaged mothers. Evidence from three United States randomized controlled trials (RCTs) on the effectiveness of this intervention at improving pregnancy outcomes, improving child health and development, and increasing maternal economic self-sufficiency is robust. However, the effectiveness of the NFP in Canada, with its different health and social care context, needs to be determined. The purpose of this article is to describe the complex process for moving the NFP from the research arena to full implementation in Canada. METHODS: This process of evaluation in Canada includes (1) adapting the intervention; (2) piloting the intervention in small-scale feasibility and acceptability studies; and (3) conducting an RCT and process evaluation through a study called the British Columbia Healthy Connections Project (BCHCP). This large-scale evaluation also creates an opportunity to expand the NFP evidence base by conducting an additional study to examine potential biological mechanisms linking intervention and behavioural outcomes in children. RESULTS: Adaptation of the NFP home-visit materials is a continuous process. A pilot project determined that it was feasible to enrol eligible women into the NFP. This pilot also determined that, in Canada, it was most appropriate for public health agencies to implement the NFP and for public health nurses to deliver the intervention. Finally, the pilot showed that this intensive home-visit program was acceptable to clients, their family members and health care providers. Through the BCHCP, the next steps - the RCT and process evaluation - are currently underway. The BCHCP will also set the foundation for long-term evaluation of key public health outcomes in a highly vulnerable population of families.

TITRE: Adaptation, mise à l'épreuve et évaluation d'interventions complexes en santé publique : leçons tirées du Nurse-Family Partnership dans le secteur de la santé publique au Canada. INTRODUCTION: Le Nurse-Family Partnership (NFP) est un programme de visites à domicile destiné aux nouvelles jeunes mères défavorisées sur le plan socioéconomique. Les données issues de trois essais contrôlés randomisés (ECR) américains ont solidement démontré l'efficacité des interventions quant à l'amélioration de l'issue de la grossesse, de la santé et du développement des enfants ainsi que de l'autonomie économique des mères. Cependant, l'efficacité du NFP dans le contexte canadien des services de santé et des services sociaux, qui diffère de celui des États-Unis, reste à déterminer. Cet article vise à décrire le processus complexe suivi pour adapter la recherche sur le NFP et mettre ainsi en oeuvre ce programme au Canada. MÉTHODOLOGIE: L'évaluation menée au Canada se divise en trois étapes : 1) adaptation de l'intervention, 2) mise à l'épreuve de l'intervention dans des études de faisabilité et d'acceptabilité à petite échelle et 3) réalisation d'un ECR et d'une évaluation du processus dans le cadre de l'étude intitulée British Columbia Healthy Connections Project (BCHCP). Cette évaluation à grande échelle permettra d'enrichir la base de données probantes du NFP par la tenue d'une étude supplémentaire sur les mécanismes biologiques susceptibles de témoigner de la relation entre l'intervention et les effets sur le comportement des enfants. RÉSULTATS: L'adaptation de la documentation du NFP pour les visites à domicile est un processus continu. Un projet pilote a montré la faisabilité du recrutement des femmes admissibles au NFP. Il a aussi révélé qu'il était préférable au Canada que le NFP soit mis en oeuvre par les organismes de santé publique et que les infirmières et infirmiers en santé publique (ISP) s'occupent des interventions. Enfin, il a montré que ce programme intensif de visites à domicile a bénéficié d'une réception positive de la part des clientes, des membres de leur famille et des fournisseurs de soins de santé. Les prochaines étapes ­ à savoir l'ECR et l'évaluation du processus ­ ont été entamées dans le cadre du BCHCP. Ce projet jettera les bases d'une évaluation à long terme des principaux résultats en matière de santé publique concernant des familles hautement vulnérables.

Ligand-dependent ubiquitination of Smad3 is regulated by casein kinase 1 gamma 2, an inhibitor of TGF-beta signaling.

Transforming growth factor-beta (TGF-beta) elicits a variety of cellular activities primarily through a signaling cascade mediated by two key transcription factors, Smad2 and Smad3. Numerous regulatory mechanisms exist to control the activity of Smad3, thereby modulating the strength and specificity of TGF-beta responses. In search for potential regulators of Smad3 through a yeast two-hybrid screen, we identified casein kinase 1 gamma 2 (CKIgamma2) as a novel Smad3-interacting protein. In mammalian cells, CKIgamma2 selectively and constitutively binds Smad3 but not Smad1, -2 or -4. Functionally, CKIgamma2 inhibits Smad3-mediated TGF-beta responses including induction of target genes and cell growth arrest, and this inhibition is dependent on CKIgamma2 kinase activity. Mechanistically, CKIgamma2 does not affect the basal levels of Smad proteins or activity of the receptors. Rather, CKIgamma2 preferentially promotes the ubiquitination and degradation of activated Smad3 through direct phosphorylation of its MH2 domain at Ser418. Importantly, mutation of Ser418 to alanine or aspartic acid causes an increase or decrease of Smad3 activity, respectively, in the presence of TGF-beta. CKIgamma2 is the first kinase known to mark activated Smad3 for destruction. Given its negative function in TGF-beta signaling and its reported overexpression in human cancers, CKIgamma2 may act as an oncoprotein during tumorigenesis.

Cost implications of introducing an alternative treatment for patients with osteoarthritis of the knee in a managed care setting.

OBJECTIVES: To illustrate the current cost of treating osteoarthritis (OA) of the knee and to demonstrate potential savings associated with the new treatment modality of viscosupplementation in a managed care setting. STUDY DESIGN: Pharmacoeconomic model with inputs obtained from peer-reviewed medical literature, clinical trial data, clinical expert opinion, and claims data. METHODS: A spreadsheet-based model was developed to define a treatment pathway for OA of the knee, illustrate the current costs of treating patients with the condition, and demonstrate the potential savings associated with introduction of Hylan G-F 20. A hypothetical cohort of patients categorized as having mild, moderate, or severe OA of the knee was followed over a 3-year time period. The analysis was conducted from the perspective of a managed care plan with a large Medicare population. RESULTS: The 3-year savings associated with adding 1 or more courses of Hylan G-F 20 therapy to the standard treatment pathway for OA of the knee was $8,810,771. The total savings per OA patient receiving Hylan G-F 20 was $4706. The number of total knee replacements (TKRs) avoided was 808. The model was highly sensitive to the durability of Hylan G-F 20; increasing and decreasing durability within a reasonable range resulted in 3-year savings of $9,131,879 and $2,012,082, respectively. CONCLUSIONS: Hylan G-F 20 has proven to be an effective treatment for patients with OA of the knee. Appropriate use of Hylan G-F 20 could delay the need for TKRs and generate savings in the managed care setting.

Effects of ionization mode on charge-site-remote and related fragmentation reactions of long-chain quaternary ammonium ions.

Comparison of collisionally activated fragment spectra of long-chain quaternary ammonium ions, formed by liquid-assisted secondary ion mass spectrometry (LSIMS) and electrospray ionization (ESI), shows the latter are dominated by radical cations while the former yield mainly even-electron charge-site-remote (CSR) fragments, similar to the report for different precursors by Cheng et al., J. Am. Soc. Mass Spectrom. 1998, 9, 840. Here, mixed-site fragmentation products (formal loss of a radical directly bonded to the nitrogen plus a radical derived from the long chain) are of comparable importance for both ionization techniques. These observations are difficult to understand if the CSR ions are formed by a concerted rearrangement-elimination reaction, since precollision internal energies of the ESI ions are much lower than those of the ions from LSIMS. Alternatively, if one discards the concerted mechanism for high-energy CA, and assumes that the even-electron fragments are predominantly formed via homolytic bond cleavage, the colder radical cations from ESI survive to the detector while the more energized counterparts from LSIMS preferentially lose a hydrogen atom to yield the CSR ions, as proposed by Wysocki and Ross (Int. J. Mass Spectrom. Ion Processes 1991, 104, 179). The present work also attempts to reconcile discrepancies involving critical energies and known structures for neutral fragments.

Maternal morbidity and mortality associated with intrauterine fetal demise: five-year experience in a tertiary referral hospital.

BACKGROUND: Risk factors for and management of intrauterine fetal demise (IUFD) have been investigated, but the maternal morbidity has not been evaluated. METHODS: Over a 60-month interval, all cases of IUFD after 20 weeks' gestation were reviewed for maternal trauma and maternal postpartum complications. RESULTS: In this retrospective analysis, 498 singleton and 24 twin pregnancies with an IUFD were identified. A cervical or perineal laceration requiring surgical repair complicated 9.4% of pregnancies. One uterine dehiscence and one uterine rupture occurred. Endometritis, the most common postpartum complication, occurred in 63 of 522 patients (12%). One maternal death occurred. Total mean hospital stay was 4.9 +/- 5.7 days. CONCLUSION: Maternal morbidity and rarely mortality can follow IUFD, but this morbidity is similar to that observed without IUFD.

The activity of guanine exchange factor NET1 is essential for transforming growth factor-beta-mediated stress fiber formation.

To examine signaling pathways underlying transforming growth factor-beta (TGF-beta)-mediated changes in cell morphology, we used a microarray system to identify downstream target genes that may play a role in this process. Through this approach, we found that the NET1 gene was induced upon TGF-beta treatment in several cell types. NET1 is a guanine nucleotide exchange factor for RhoA whose activity has been implicated in stress fiber formation. In the Swiss 3T3 cell line, TGF-beta induces NET1 expression, and this correlated with an increase in stress fiber formation. Overexpression of the wild type NET1 gene increases stress fiber formation, and overexpression of a dominant negative NET1 mutant (L392E) prevented TGF-beta dependent increase in stress fiber formation. Furthermore, treatment of the cells with a RhoA kinase inhibitor Y-27632 blocks TGF-beta-induced stress fiber formation. By using a stable cell line expressing dominant negative Smad3, we found that the Smad signaling pathway is essential for the induction of NET1, which in turn leads to the increase of Rho activity. Taken together, those data suggest that induction of NET1 is important for the increase of Rho activity upon TGF-beta treatment, which may represent the critical trigger for a variety of downstream events in different cells. Our results support the presence of a novel signaling pathway by which TGF-beta may regulate the formation of stress fibers and reorganization of cytoskeletal structures.

Measurement issues in promoting continued competence.

PURPOSE: Promoting continued competence in nursing practice has always been of concern to staff development and continuing education educators. Recently it has reached new levels of significance. As a result of the proposals made by the Pew Commission Taskforce on Health Care Workforce Regulation and the National Council of State Boards of Nursing, the profession is re-examining the question, "How do we promote and assure continued competence?" The purpose of this article is to examine the measurement issues related to answering this question. FINDINGS: The nursing literature provides several definitions of competence, but none are operational. To measure a concept it must be operationalized. Mandatory continuing education, peer review, and practice or process audits have been used to assess continued competence. The measurement issues involved in assessing competence include selecting an appropriate measurement paradigm, selecting accurate measurement instruments, and interpreting the measurement data. IMPLICATIONS: Staff development educators and nurse managers should collaborate in the establishment of standards of practice, the identification of essential competencies specific to the setting, and the development of appropriate methods of measurement of these competencies.

Low levels of cadmium chloride alter the immunoprecipitation of corneal cadherin-complex proteins.

The effect of cadmium chloride on the immunoprecipitation of cadherin and the associated adherens junctional proteins, alpha- and beta-catenin, was examined in isolated bullfrog (Rana catesbeiana) corneas utilizing Western blot and enhanced chemoluminescent techniques. Application of either 1.0 microM or 75.0 microM CdCl2 to the corneal endothelium for 2 h markedly decreased the immunoprecipitation of cadherins as compared to paired control corneas. Immunoprecipitation of alpha-catenin was increased in response to both doses of CdCl2, while the immunoprecipitation of beta-catenin was little changed by either cadmium dose. There is accumulating evidence that cadmium may increase epithelial paracellular permeability by interfering with cadherin complex activity at intercellular junctions. The present study suggests that inorganic cadmium in low micromolar concentrations may decrease the integrity of the corneal endothelium, at least in part through a similar mechanism involving disruption of junctional cadherin complex function.

Use of learning contracts in a RN-to-BSN leadership course.

Educating RN-to-BSN students can be challenging and rewarding. The diversity of these students can make designing courses quite difficult. In addition, the RN with experience in nursing practice may resent being put in the traditional role of student. Nominal group process and use of learning contracts in a leadership course in one RN completion program are described as two means of meeting the unique learning preferences of working, adult learners. Houle (1984) identified the learning contract as one means of assisting the adult learner seeking continuing professional education. Accordingly, examples of use of these two strategies in continuing nursing education and staff development are described.

An evaluation of international nursing education using interactive desktop video conferencing.

Evolving computer technology has opened the door to unique and previously unavailable educational experiences. The use of one form of distance learning enabled undergraduate nursing students in the United States and England to participate in a short course on women's health issues. The nursing faculty at two universities, one in the United States and the other in England, collaborated on an international nursing education project. The delivery device was interactive desktop video conferencing (DTV). The focus of this article is on the evaluation of the project with recommendations about how best to use DTV in international nursing education.

Systematic planning of continuing nursing education.

To avoid omissions and to promote efficient use of time, it is important to plan continuing nursing education (CNE) activities systematically. A detailed and comprehensive planning checklist is provided to assist the novice planner to organize and implement a CNE offering competently.

Genetic engineering for high methionine grain legumes.

Methionine (Met) is the primary limiting essential amino acid in grain legumes. The imbalance in amino acid composition restricts their biological value (BV) to 55 to 75% of that of animal protein. So far improvement of the BV could not be achieved by conventional breeding. Therefore, genetic engineering was employed by several laboratories to resolve the problem. Three strategies have been followed. A) Engineering for increased free Met levels; B) engineering of endogenous storage proteins with increased numbers of Met residues; C) transfer of foreign genes encoding Met-rich proteins, e.g. the Brazil nut 2S albumin (BNA) and its homologue from sunflower, into grain legumes. The latter strategy turned out to be most promising. In all cases the gene was put under the control of a developmentally regulated seed specific promoter and transferred into grain legumes using the bacterial Agrobacterium tumefaciens-system. Integration into and copy numbers in the plant genome as well as Mendelian inheritance and gene dosage effects were verified. After correct precursor processing the mature 2S albumin was intracellularly deposited in protein bodies which are part of the vacuolar compartment. The foreign protein amounted to 5 to 10% of the total seed protein in the best transgenic lines of narbon bean (Vicia narbonensis L., used in the authors' laboratories), lupins (Lupinus angustifolius L., used in CSIRO, Australia), and soybean (Glycine max (L.) Merr., used by Pioneer Hi-Bred, Inc., USA). In the narbon bean the increase of Met was directly related to the amount of 2S albumin in the transgenic seeds, but in soybean it remained below the theoretically expected value. Nevertheless, trangenic soybean reached 100%, whereas narbon bean and lupins reached approximately 80% of the FAO-standard for nutritionally balanced food proteins. These results document that the Met problem of grain legumes can be resolved by genetic engineering.

Saquinavir, an HIV protease inhibitor, is transported by P-glycoprotein.

Saquinavir, a peptidomimetic HIV protease inhibitor, has been shown to be effective in reducing patient viral load and reducing mortality. In this report we investigated whether saquinavir is a substrate for the multidrug resistance transporter P-glycoprotein (P-gp), which may reduce the effective intracellular concentration of the drug. G185 cells, which highly express P-gp, are resistant to saquinavir-mediated cytotoxicity, and co-administration of cyclosporine reversed this resistance. Saquinavir and saquinavir mesylate inhibited basolateral to apical transport of the fluorescent dye rhodamine 123 in a polarized epithelial transport assay, a result that suggests competition of these drugs for the P-gp transporter. Finally, we measured specific, directional transport of saquinavir and saquinavir mesylate in an epithelial monolayer model. Transport in the basolateral to apical direction was 3-fold greater than apical to basolateral flux for both saquinavir and saquinavir mesylate and was blocked by co-incubation with the established P-gp reversal agents cyclosporine and verapamil. These data provide evidence that saquinavir is a substrate for the P-gp transporter and suggest that this protein may affect intracellular accumulation of the drug and contribute to its poor oral bioavailability.