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Optimized Hot Phenol-Based RNA Extraction from Mycobacteria: A Robust Approach for Reliable Gene Expression Analysis.

Mycobacterium tuberculosis (Mtb) remains a major global health threat, underscoring the need for reliable transcriptomic studies to understand its biology and drug resistance mechanisms. Such analyses depend on obtaining high-quality, high-yield RNA. Although several RNA extraction methods are available, many require expensive reagents, large culture volumes, or specialized equipment, limiting their suitability for large-scale studies, particularly in resource-constrained settings. Here, an optimized Hot Phenol based RNA extraction method specifically tailored for mycobacteria is presented. The method uses minimal culture volume and commonly available reagents to consistently yield high-quality RNA suitable for high-throughput transcriptomic applications. RNA quantity and integrity were assessed by gel electrophoresis and RNA integrity analysis (RIN), and its suitability for downstream applications was confirmed by qPCR and Qubit 4. To benchmark the performance of the optimized method, a parallel RNA extraction using TRIzol and RNeasy under identical experimental conditions was carried out, including the same Mycobacterium species, culture volume, growth phase (logarithmic and stationary), and lysis conditions. This allowed a direct comparison of yield, quality, feasibility, and cost. The optimized Hot Phenol method demonstrated comparable or improved RNA yield and quality while significantly reducing reagent cost and dependence on specialized equipment. Owing to its efficiency, reproducibility, and affordability, this protocol provides a practical alternative for large-scale gene expression and transcriptomic studies in Mtb and other mycobacterial species.

RNA, Bacterial↗

Panel discussion. Data needs in cancer.

A prospective, comprehensive outcomes database was recently initiated by the National Comprehensive Cancer Network (NCCN) after a 2-year study to test data collection methods and systems. It started with data on 400 patients with newly diagnosed breast cancer at five NCCN sites, and over the next 3 years is projected to grow to include more than 12,000 patients with common cancers treated at all eligible NCCN sites. Among the goals of the database are: 1) to establish the capability to select, analyze, and report patterns of care and outcomes; 2) to allow NCCN members to assess their compliance with NCCN clinical practice guidelines and benchmark their performance against the rest of the NCCN; 3) to establish a true databased continuous quality improvement program; 4) to support clinical disease-oriented research and methodologic studies; and 5) to provide the NCCN with a vehicle for forging partnerships with others in the health-care field, such as the pharmaceutical industry, regulatory agencies, and accrediting bodies. Many of those potential partners were represented on this panel. Panelists discussed the data needs of their organizations, what they are doing to meet those needs, and how a comprehensive database will ultimately help improve patient care.

Aged↗

Broader range of skills distinguishes successful CFOs.

In recent years, healthcare CFOs have seen their role expand significantly beyond traditional financial duties. A series of trended surveys on CFO roles and responsibilities reveals that today's healthcare CFO requires a broad new range of traits and skills in the areas of leadership, operations, and healthcare strategy. CFOs regard strategic thinking and the ability to communicate clearly as the most important of their essential leadership traits and skills, respectively. Among operational and strategic skills, CFOs most often cite the importance of being able to improve organizational performance and benchmark. Healthcare CFOs can enhance their chances of success by focusing self-improvement efforts on five key areas: implementing the organization's vision; developing tactics that stimulate change; enhancing communication skills; focusing on managing and leading; and strengthening relationships.

Benchmarking↗

Demonstrating quality at the physician-office level.

A multidisciplinary task force consisting of administration, quality management, directors of service, and network development staff of a nonprofit organization developed and implemented a comprehensive report card process for its 50 physician-office sites. The goals were to develop and implement a process that would provide a comprehensive summary of the administrative and clinical activity of all network sites, to develop a framework for identifying best practices within the network, and to enhance motivation to improve performance through benchmarking with physician offices within the network. This initiative also sought to transform the voluminous data collected into meaningful information that could be used to improve the process of operations and outcomes.

Benchmarking↗

[External quality assessment of clinical services in Europe].

All countries and clinical specialties have some elements of systems for quality improvement, but their aims, configurations, models standards and assessments are often not formally recognised or integrated. External programmes to assess service delivery in Europe include the International Standardisation Organisation (ISO) and Excellence (EFQM - European Foundation Quality Management) models (industry based, management focus), peer review and accreditation (health care based, professional focus) and inspection (regulatory, safety-focused). Patient surveys and disease registers also contribute to assessment and benchmarking best performance. There are legal, cultural, professional and commercial reasons to adopt common core standards but there is little legislative framework to allow formal harmonisation within and between countries and clinical specialties. Examples are given of various approaches to the definition, assessment and improvement of standards for clinical services in Europe in order to encourage specialist associations to develop self-regulations based on the experience of others. Clinical practice and clinical services could be more efficiently and effectively harmonised by the professions than by their respective governments.

Accreditation↗

Off the shelf or recalibrate? customizing a risk index for assessing mortality.

BACKGROUND: Public "report cards" for cardiac surgery have been freely available from a variety of sources. These risk-adjusted indices serve as a means of benchmarking outcomes performances, allowing comparisons of outcomes between surgical programs, and quantifying quality improvement programs. We examined two alternative strategies for using previously developed risk-adjusted mortality models in a community hospital: (1) using the model "off the shelf" (OTS) and (2) recalibrating the existing model (RM) to fit the institution-specific population. METHODS: Six OTS models were used: Parsonnet (PA), Canadian (CA), Cleveland (CL), Northern New England (NNE), New York (NY), and New Jersey (NJ). The RM models were created by each model's independent variables and definitions and adjusting the weighting with logistic regression methods. The accuracy, the C statistic, and the precision of each model were assessed for in-hospital mortality. We compared the OTS version of each model to the RM version with methods detailed by Hanley and McNeil. RESULTS: The RM C statistic was improved for all risk-adjusted models, most notably in the statistical improvement seen in the PA (0.053 improvement) and NJ (0.052 improvement) indices. Statistical gains in precision were also seen in the RM models for the PA, CL, and NNE indices. Conversely, one model, the CA model, was more poorly calibrated in the RM model compared with the OTS model, despite an improved C statistic (0.062). CONCLUSIONS: The RM strategy provides institution-explicit models that demonstrate a higher degree of accuracy and precision than the OTS models.

Age Factors↗

Outsourcing in the public refuse collection sector: exploiting old certainties or exploring new possibilities?

An outsourcing process in a medium-size city, by Norwegian standards, provided the background for our study. In 1996, the city council decided to contract out the refuse collection in half of its total area (the western part of the city), and to let the public refuse collection enterprise continue to collect in the other half of the area. The public enterprise also participated in the competitive tender, without success, however. The bid submitted by the public refuse collection enterprise was used as the basis for a benchmarking process performed by the chief administrative officer of the city in May 1999. The process resulted in the requirement that the public enterprise downsize its number of refuse collectors by 27 per cent. The city thus acquired a more cost-effective refuse collection system both in the western part, now run by a private company, and in the rest of the city, where the collection remained the responsibility of the public enterprise, in its newly pared-down and reorganized version. Detectable changes in the refuse collectors' health status in the three-year period immediately following the outsourcing constitute our main focus in this study. We found that the downsizing and reorganization of the public enterprise had a negative impact on the refuse collectors' health status. One year after the downsizing, six of the 27 refuse collectors remaining in the public enterprise had been diagnosed with heart problems or musculoskeletal pains that were sufficiently serious to form the basis of their individual approaches to secure a disability pension, which all six were in the process of acquiring. The registered sick leave among the refuse collectors showed a dramatic increase during the same period. We found clear indications that the refuse collectors remaining in the employment of the public enterprise had no alternatives in the labour market. Although not the focus of this study, we found that the totality of costs and benefits, not only for the enterprise in question, but also for local and national authorities, needs to be considered in any realistic assessment of the impact of public sector outsourcing of activities.

Humans↗

The ICCD benchmarks for clubhouses: a practical approach to quality improvement in psychiatric rehabilitation.

OBJECTIVE: The study evaluated whether the average performance of clubhouses certified by the International Center for Clubhouse Development (ICCD) should be considered valid benchmarks for clubhouse programs. METHODS: A representative sample of clubhouses more than three years old that were based on the Fountain House model participated in a 1998 mail survey. To verify that ICCD certification is a valid indicator of program quality for use in setting benchmark performance rates, 71 certified and 48 noncertified programs were compared on a variety of organizational variables. RESULTS: Even though certified and noncertified clubhouses were similar in organizational structure and resources, findings from a logistic regression analysis confirmed that certified clubhouses provided a wider array of rehabilitation services and achieved higher rates of employment. CONCLUSIONS: The findings suggest that ICCD certification is a valid indicator of program quality. The ICCD has therefore proposed that the average performance of certified U.S. clubhouses in specific domains be adopted as benchmarks for organizational performance. When tailored for programs in particular regions and with specific levels of funding, the ICCD benchmarks for clubhouse performance set fair and reasonable expectations for clubhouse programs and for the design of performance contracts between departments of mental health and ICCD clubhouses.

Benchmarking↗

Determination of an occupational exposure guideline for manganese using the benchmark method.

An occupational risk assessment for manganese (Mn) was performed based on benchmark dose analysis of data from two epidemiological studies providing dose-response information regarding the potential neurological effects of exposure to airborne Mn below the current Occupational Safety and Health Administration (OSHA) Permissible Exposure Level (PEL) of 5 mg Mn/m3. Based on a review of the scientific evidence regarding the toxicity of Mn, it was determined that the most appropriate measure of exposure to airborne Mn for the subclinical effects measured in these studies is recent (rather than historical or cumulative) concentration of Mn in respirable (rather than total) particulate. For each of the studies analyzed, the individual exposure and response data from the original study had been made available by the investigators. From these two studies benchmark concentrations calculated for eight endpoints ranged from 0.09 to 0.27 mg Mn/m3. From our evaluation of these results, and considering the fact that the subtle, subclinical effects represented by the neurological endpoints tested in these studies do not represent material impairment, we believe an appropriate occupational exposure guideline for manganese would be in the range of 0.1 to 0.3 mg Mn/m3, based on the respirable particulate fraction only, and expressed as an 8-hour time-weighted average.

Air Pollutants, Occupational↗

Better information for the board.

Healthcare managers are making quicker, riskier decisions in an increasingly competitive and regulated environment. Questions have been raised regarding the accountability and performance of boards of these organizations, as board members are not always selected based on their competencies to guide such decisions. Adapting mission and strategy and monitoring organizational performance require information that boards get mostly from management. The purpose of this study was to examine the information that boards regularly get to carry out their functions. I obtained board documents from four not-for-profit hospitals and health systems in different boroughs of New York City. At each institution, I conducted one-hour interviews with at least three board members and three top managers. I also attended at least one board or executive committee meeting and one additional meeting, usually of the finance committee. Principal findings were that the boards get too much data, the same data that management gets, and little comparative data on performance of similar benchmarked organizations. Board members and managers are satisfied with the information that board members get and have no plans to improve their system of shaping, or the quality of, information. Key recommendations to boards and managers are: (1) boards must take greater responsibility for identifying the information that they get and how they wish to get it, (2) managers must ensure that measurable objectives are developed, against which organizational performance can be evaluated, (3) boards must get information that is targeted and shaped to better fit board functions, (4) managers must develop information sets for main service lines, (5) boards must get information on the expectations and satisfaction levels of key stakeholders, (6) boards must get better and more focused information on performance of benchmarked institutions, and (7) boards must get less hospital operating data on a monthly basis.

Decision Making, Organizational↗

Benchmarking hospital laboratory financial and operational performance.

The movement toward more integrated delivery systems requires hospital administrators, medical staffs, and health care network organizations to consider strategies that will meet the future challenges facing laboratory services. Many health care experts predict that the number of hospital inpatient days, staffed acute care beds, and length of stay will continue their precipitous decline, and then stabilize during the next four to five years. Hospitals should carefully evaluate how their laboratories might be affected as a result of the decline in inpatient services and the integration of health care services at all levels. Hospital executive management must find a way to manage staffing levels and technical resources in order to maintain quality patient services in the face of declining test volume. This Special Report discusses relevant benchmarks intended to help hospital administrators and laboratory directors identify "best practices" in hospital laboratories so that comparisons of patterns of care and financial operations can be made. Benchmarking the relative financial and operational performance of hospital laboratories allows health care planners to design the most appropriate laboratory services delivery system for future hospital inpatient and outpatient market demands. Factors influencing financial and operation performance will be investigated, including utilization, testing costs, staffing mix, productivity, and organizational structure. This will be followed by a discussion on the future of laboratories and the trend toward regional laboratories owned by hospital consortiums.

Clinical Laboratory Techniques↗

Extension of the IWA/COST simulation benchmark to include expert reasoning for system performance evaluation.

In this paper the development of an extension module to the IWA/COST simulation benchmark to include expert reasoning is presented. This module enables the detection of suitable conditions for the development of settling problems of biological origin (filamentous bulking, foaming and rising sludge) when applying activated sludge control strategies to the simulation benchmark. Firstly, a flow diagram is proposed for each settling problem, and secondly, the outcome of its application is shown. Results of the benchmark for two evaluated control strategies illustrate that, once applied to the simulation outputs, this module provides supplementary criteria for plant performance assessment. Therefore, simulated control strategies can be evaluated in a more realistic framework, and results can be recognised as more realistic and satisfactory from the point of view of operators and real facilities.

Benchmarking↗

Why use noise?

Measuring the dependence of visual sensitivity on parameters of the visual stimulus is a mainstay of vision science. However, it is not widely appreciated that visual sensitivity is a product of two factors that are each invariant with respect to many properties of the stimulus and task. By estimating these two factors, one can isolate visual processes more easily than by using sensitivity measures alone. The underlying idea is that noise limits all forms of communication, including vision. As an empirical matter, it is often useful to measure the human observer's threshold with and without a noise background added to the display, to disentangle the observer's ability from the observer's intrinsic noise. And when we know how much noise there is, it is often useful to calculate ideal performance of the task at hand, as a benchmark for human performance. This strips away the intrinsic difficulty of the task to reveal a pure measure of human ability. Here we show how to do the factoring of sensitivity into efficiency and equivalent noise, and we document the invariances of the two factors.

Artifacts↗

Study documents wide-ranging efficiency in knee arthroscopies.

Data Benchmarks: How long does it take you to perform a knee arthroscopy with meniscectomy? A new study takes a look at how fast 46 surgery centers prepare for, perform and turn around operating rooms for the common outpatient procedure. You'll be surprised at how widely varied performance is.

Arthroscopy↗

Systematic tuning of parameters in support vector clustering.

Clustering algorithms divide a set of observations into groups so that members of the same group share common features. In most of the algorithms, tunable parameters are set arbitrarily or by trial and error, resulting in less than optimal clustering. This paper presents a global optimization strategy for the systematic and optimal selection of parameter values associated with a clustering method. In the process, a performance criterion for the optimization model is proposed and benchmarked against popular performance criteria from the literature (namely, the Silhouette coefficient, Dunn's index, and Davies-Bouldin index). The tuning strategy is illustrated using the support vector clustering (SVC) algorithm and simulated annealing. In order to reduce the computational burden, the paper also proposes an alternative to the adjacency matrix method (used for the assignment of cluster labels), namely the contour plotting approach. Datasets tested include the iris and the thyroid datasets from the UCI repository, as well as lymphoma and breast cancer data. The optimal tuning parameters are determined efficiently, while the contour plotting approach leads to significant reductions in computational effort (CPU time) especially for large datasets. The performance criteria comparisons indicate mixed results. Specifically, the Silhouette coefficient and the Davies-Bouldin index perform better, while the Dunn's index is worse on average than the proposed performance index.

Algorithms↗

Benchmarking in home care.

Recognizing that continuous quality improvement will be imperative for success in the future, six visiting nurse associations of northwestern Pennsylvania used benchmarking as a tool to improve their respective agencies' performance. The agencies benchmarked three areas: continuity of care, patient rehospitalization, and nosocomial infection rate.

Continuity of Patient Care↗