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Multivariate statistical tools applied to the characterization of the proteomic profiles of two human lymphoma cell lines by two-dimensional gel electrophoresis.

Mantle cell lymphoma (MCL) cell lines have been difficult to generate, since only few have been described so far and even fewer have been thoroughly characterized. Among them, there is only one cell line, called GRANTA-519, which is well established and universally adopted for most lymphoma studies. We succeeded in establishing a new MCL cell line, called MAVER-1, from a leukemic MCL, and performed a thorough phenotypical, cytogenetical and molecular characterization of the cell line. In the present report, the phenotypic expression of GRANTA-519 and MAVER-1 cell lines has been compared and evaluated by a proteomic approach, exploiting 2-D map analysis. By univariate statistical analysis (Student's t-test, as commonly used in most commercial software packages), most of the protein spots were found to be identical between the two cell lines. Thirty spots were found to be unique for the GRANTA-519, whereas another 11 polypeptides appeared to be expressed only by the MAVER-1 cell line. A number of these spots could be identified by MS. These data were confirmed and expanded by multivariate statistical tools (principal component analysis and soft-independent model of class analogy) that allowed identification of a larger number of differently expressed spots. Multivariate statistical tools have the advantage of reducing the risk of false positives and of identifying spots that are significantly altered in terms of correlated expression rather than absolute expression values. It is thus suggested that, in future work in differential proteomic profiling, both univariate and multivariate statistical tools should be adopted.

Cell Line, Tumor↗

The revision of a workload measurement tool to reflect the nursing needs of patients with traumatic brain injury.

Rehabilitation for persons with head injuries is a multifaceted process that requires structure, consistency, and predictability. This article discusses the process followed by nurses on one rehabilitation unit to revise a standard nursing workload measurement tool to reflect the physical, cognitive, behavioral, and psychosocial needs of patients with traumatic brain injury (TBI). Family education and support, a critical component of care, was also addressed. The flexibility of the tool, known as GRASP, allowed the incorporation of neurobehavioral items based on the Rancho Los Amigos cognitive functioning scale and the inclusion of important therapeutic nursing interventions. The revised tool reflects the time required to deliver comprehensive nursing care to TBI patients and, thus, promotes the desired rehabilitation outcomes for patients and their families.

Brain Injuries↗

The H-reflex as a tool in neurophysiology: its limitations and uses in understanding nervous system function.

The Hoffmann reflex (H-reflex) is extensively used as both a research and clinical tool. The ease with which this reflex can be elicited in several muscles throughout the body makes it an attractive tool. This review discusses some of the important limitations in using the H-reflex. In particular, the inaccurate but widely held assumptions that the H-reflex (1). represents the monosynaptic reflex of the Ia afferent onto homonymous motoneurons, and (2). can be used to measure motoneuronal excitability are addressed. The second part of this review explores the utility of the H-reflex as a neural probe in neurophysiology and motor control research. Applications ranging from the investigation of the functional organization of neural circuitry to the study of adaptive plasticity in spinal structures in health and disease suggest that the H-reflex will continue to be an extensively used tool in motor control neurophysiology.

Animals↗

The pitfalls of proteomics experiments without the correct use of bioinformatics tools.

The elucidation of the entire genomic sequence of various organisms, from viruses to complex metazoans, most recently man, is undoubtedly the greatest triumph of molecular biology since the discovery of the DNA double helix. Over the past two decades, the focus of molecular biology has gradually moved from genomes to proteomes, the intention being to discover the functions of the genes themselves. The postgenomic era stimulated the development of new techniques (e.g. 2-DE and MS) and bioinformatics tools to identify the functions, reactions, interactions and location of the gene products in tissues and/or cells of living organisms. Both 2-DE and MS have been very successfully employed to identify proteins involved in biological phenomena (e.g. immunity, cancer, host-parasite interactions, etc.), although recently, several papers have emphasised the pitfalls of 2-DE experiments, especially in relation to experimental design, poor statistical treatment and the high rate of 'false positive' results with regard to protein identification. In the light of these perceived problems, we review the advantages and misuses of bioinformatics tools - from realisation of 2-DE gels to the identification of candidate protein spots - and suggest some useful avenues to improve the quality of 2-DE experiments. In addition, we present key steps which, in our view, need to be to taken into consideration during such analyses. Lastly, we present novel biological entities named 'interactomes', and the bioinformatics tools developed to analyse the large protein-protein interaction networks they form, along with several new perspectives of the field.

Amino Acid Sequence↗

The potential use of continuous-flow isotope-ratio mass spectrometry as a tool in forensic soil analysis: a preliminary report.

The use of continuous-flow isotope-ratio mass spectrometry (CF-IRMS) as a tool in soil analysis has been assessed as part of a larger study using a number of geological techniques applied in a forensic context. Carbon and nitrogen isotopic ratios, delta13C and delta15N, have been analysed to investigate situations which have arisen from crime casework. Three questions have been addressed: the role of spatial variation found over the short-scale (less than 20 m), temporal variation over a period of almost 2 years, and the variation found between source soils and soil transferred to footwear soles during a simple one-stage transfer process. Results are presented for the three experiments. The use of carbon and nitrogen isotopes has been shown to be useful in discriminating between soil types and sample locations, even when sampling occurs at a different time (as might be the case with a crime scene). In cases of primary transfer (from a source soil by a one-stage transfer to another surface, in this case, shoes and boots), the combination of carbon and nitrogen isotope ratios is a valuable tool in discriminating between sites and in showing the relationship of the transferred samples to the relevant source soils. Used in combination with other analytical techniques, isotopic analysis may prove to be a useful tool in a forensic context.

Carbon↗

"Derived spectra" software tools for detecting spatial and spectral features in spectrum images.

A spectrum image is recorded as an x-y array of beam locations at each of which a spectrum of radiation is recorded as stimulated by the beam. The large database or "datacube" that results from a single image presents a significant challenge to the analyst to recover information efficiently, especially in the case where a true unknown is examined. This paper describes a class of "derived spectra" software tools that can aid the analyst in recognizing both common and rare features within the datacube. A derived spectrum tool creates a spectrum-like display (intensity vs. channel number) in which the intensity (e.g., x-ray counts) at a particular channel (e.g., x-ray photon energy) is calculated from all of or a subset of the pixel intensities measured for that channel. Derived spectra tools considered include the SUM, MAXIMUM PIXEL, RUNNING SUM, and RUNNING MAXIMUM. The SUM-derived spectrum is useful for recognizing common features of the datacube, while the MAXIMUM PIXEL-and RUNNING MAXIMUM-derived spectra can locate rare, unanticipated features, which may occur as infrequently as being present at a single pixel in the original datacube.

Journal Article↗

Hand function and tool behavior in early hominids.

Prompted by the discovery of the Olduvai Hominid 7 hand (Homo habilis) in 1960, studies of primate hands were conducted in order to understand better the functional morphology of the earliest hominids and the evolution of tool behavior. Since Napier's work in the early 1960's, the hand bones of a number of other species have been recovered in East and South Africa. New hominid finds and a prolific archaeological record have broadened our perspective on the evolution of tool behavior in Australopithecus, Paranthropus, and early Homo. A debate is underway at this time on the questions of what exactly we can glean from the fossil record regarding the evolution of tool behavior and what are the best approaches to interpreting the evidence. I support the model of Napier that identified morphological correlates of precision and power grasping in the hands of extant primates and in early hominid hand bones. At the same time, I question both the underlying rationale and attempts to identify more subtle aspects of precision grasping, based on present evidence.

Animals↗

Modified phage peptide libraries as a tool to study specificity of phosphorylation and recognition of tyrosine containing peptides.

Tyrosine phosphorylation and protein recognition, mediated by phosphotyrosine containing peptides, play an important role in determining the specific response of a cell, when stimulated by external signals. We have used peptide repertoires displayed by filamentous phage as a tool to study the substrate specificity of the protein tyrosine kinase (PTK) p55(fyn) (Fyn). Peptide libraries were incubated for a short time in the presence of Fyn and phages displaying efficiently phosphorylated peptides were selected by panning over anti-phosphotyrosine antibodies. The characterization of the peptides enriched after three phosphorylation/selection rounds allowed us to define a canonical substrate sequence for the kinase Fyn, E-(phi/T)YGx phi, where phi represents any hydrophobic residue. A peptide conforming to this sequence is a better substrate than a second peptide designed to be in accord with the consensus sequence recognised by the Fyn SH2 domain. When the library phosphorylation reaction is carried out in saturation conditions, practically all the tyrosine containing peptides are phosphorylated, irrespective of their context. These "fully modified" peptide libraries are a valuable tool to study the specificity of phosphotyrosine mediated protein recognition. We have used this new tool to identify a family of peptides that bind the PTB domain of the adapter protein Shc. Comparison of the peptide sequences permits us to confirm the essential role of N at position -3, while P often found at position -2 in natural targets is not absolutely required. Furthermore, our approach permits us to reveal an "extended" consensus indicating that residues that do not seem to influence binding in natural peptides can make productive contacts, at least in linear peptides.

Consensus Sequence↗

Surrogate genetics: the use of bacterial hybrids as a genetic tool.

Experimental dissection of bacterial genomes requires a well-developed set of genetic tools, but many bacteria lack the essential tools required for genetic analysis. Recombination of a region of chromosomal DNA from poorly characterized donor bacteria with the chromosome of a suitable surrogate host creates a genetically malleable hybrid, providing a short-cut for the detailed genetic analysis of the substituted genes. However, recombination between closely related but nonidentical DNA sequences ("homeologous recombination") is strongly inhibited, posing a powerful barrier to gene exchange between bacteria and a major impediment to the construction of genetic hybrids. By taking advantage of mutS and recD mutant recipients, it is possible to effectively overcome the recombination barrier, allowing construction of genetic hybrids in a related surrogate host. Once stably recombined into the recipient chromosome, the donor DNA can be studied with all the genetic tools available in the surrogate host. In addition to facilitating standard genetic analysis, use of a surrogate host can provide novel approaches to study the physiological roles of unique genes from poorly characterized bacteria.

Bacteria↗

Electromagnetic field focusing (EFF) probe--a new neurosurgical tool.

The electromagnetic field focusing (EFF) probe is capable of producing well circumscribed, intense heat at the point of contact with the tissue. Experimental studies were carried out to assess this probe as a neurosurgical tool using 38 rats and 4 mice with mammary carcinoma. The study on the rats included study of the cutting, coagulating and vaporizing effect on brain tissue including study of blood brain barrier disruption and heat dissipation. The study on the mice included the study of vaporizing property of the probe on solid tumours. The probe proved to be an excellent tool for cutting, coagulating and vaporizing purposes with very minimal disruption of blood brain barrier and demonstrated well circumscribed heating pattern. The study indicates that this tool combines the beneficial effect of both the YAG and CO2 laser.

Animals↗

A new application tool for the ROCHE pH electrode.

Using this tool the application of the pH electrode is nearly as easy as the attachment of a common ECG-scalp-electrode [3]. Visual control and lithotomy position is not necessary any more. The different tools are adapted to each other so that incision place and the tip of the pH electrode are automatically centered perpendicularly. The first results by using this tool promise to be successful. Time of application is short, the maintenance of the pH electrode is good and the application for the patient no longer disagreeable.

Electrodes↗

Tool-assisted mesh generation based on a tissue-growth model.

An heuristic mesh generation technique is proposed that is based on the model of forced particle motion, an edgewise cell-splitting algorithm and a moving tool concept. The method differs from conventional mesh generators in that it uses outward growth of the mesh, in contrast to the inward growth used in traditional meshing techniques. The method does not require prior meshing and patching of two-dimensional (2D) boundary surfaces. Instead, it uses a pre-defined skeleton of one-dimensional segments, or an arbitrary tool motion in three-dimensional (3D) space. In this respect, the technique can be considered as a 3D extension of a 2D drawing tool and can find applications in virtual reality systems. The method also guarantees the smoothness of the outer boundary of the mesh at each step of mesh generation, which is not the case with traditional propagating-front methods. The approach is based on the model of tissue growth and is suitable for meshing complex networks of bifurcating branches commonly found in biological structures: blood vessels, lungs, neural networks, plants etc. The generated meshes were used in solving unsteady flow and particle transport problems in lungs.

Biological Products↗

Architectural design and tools to support the transparent access to hospital information systems, radiology information systems, and picture archiving and communication systems.

The fragmentation of the electronic patient record among hospital information systems (HIS), radiology information systems (RIS), and picture archiving and communication systems (PACS) makes the viewing of the complete medical patient record inconvenient. The purpose of this report is to describe the system architecture, development tools, and implementation issues related to providing transparent access to HIS, RIS, and PACS information. A client-mediator-server architecture was implemented to facilitate the gathering and visualization of electronic medical records from these independent heterogeneous information systems. The architecture features intelligent data access agents, run-time determination of data access strategies, and an active patient cache. The development and management of the agents were facilitated by data integration CASE (computer-assisted software engineering) tools. HIS, RIS, and PACS data access and translation agents were successfully developed. All pathology, radiology, medical, laboratory, admissions, and radiology reports for a patient are available for review from a single integrated workstation interface. A data caching system provides fast access to active patient data. New network architectures are evolving that support the integration of heterogeneous software subsystems. Commercial tools are available to assist in the integration procedure.

Computer Systems↗

Design and implementation of World Wide Web-based tools for image management in computed tomography, magnetic resonance imaging, and ultrasonography.

This article describes our experience in developing and using several web-based tools to facilitate access to and management of images from inside and outside of our department. Having recently eliminated film in ultrasound, computed tomography (CT) and magnetic resonance imaging (MRI), a simple method was required to access imaging from computers already existing throughout the hospital. The success of the World Wide Web (WWW), the familiarity of endusers with web browsers, and the relative ease of developing user interfaces virtually dictated that such an approach be pursued in our case. The resulting web-based tools allow validated users to search our Digital Imaging and Communications in Medicine (DICOM)-compliant archive servers for specific exams; to download image data from a remote site; to request the retrieval of data from long-term storage; to view images, and to perform certain DICOM routing operations. The existing infrastructure of the internet has allowed us to develop a low-cost system capable of being used for teleradiology. Since low-level, machine-specific interface programming was avoided, these tools were developed rapidly and are easily adapted. The familiarity of browser-based interfaces has facilitated user acceptance, and the benefit of platform independence minimizes software portability concerns.

Computer Communication Networks↗

Assessment of cytochrome P450 sequences offers a useful tool for determining genetic diversity in higher plant species.

To investigate and develop new genetic tools for assessing genome-wide diversity in higher plant-species, polymorphisms of gene analogues of mammalian cytochrome P450 mono-oxygenases were studied. Data mining on Arabidopsis thaliana indicated that a small number of primer-sets derived from P450 genes could provide universal tools for the assessment of genome-wide genetic diversity in diverse plant species that do not have relevant genetic markers, or for which, there is no prior inheritance knowledge of inheritance traits. Results from PCR amplification of 51 plant species from 28 taxonomic families using P450 gene-primer sets suggested that there were at least several mammalian P450 gene mammalian-analogues in plants. Intra- and inter- specific variations were demonstrated following PCR amplifications of P450 analogue fragments, and this suggested that these would be effective genetic markers for the assessment of genetic diversity in plants. In addition, BLAST search analysis revealed that these amplified fragments possessed homologies to other genes and proteins in different plant varieties. We conclude that the sequence diversity of P450 gene-analogues in different plant species reflects the diversity of functional regions in the plant genome and is therefore an effective tool in functional genomic studies of plants.

Arabidopsis↗

Bedside waveforms interpretation as a tool to identify patient-ventilator asynchronies.

OBJECTIVE: During assisted modes of ventilatory support the ventilatory output is the final expression of the interaction between the ventilator and the patient's controller of breathing. This interaction may lead to patient-ventilator asynchrony, preventing the ventilator from achieving its goals, and may cause patient harm. Flow, volume, and airway pressure signals are significantly affected by patient-ventilator interaction and may serve as a tool to guide the physician to take the appropriate action to improve the synchrony between patient and ventilator. This review discusses the basic waveforms during assisted mechanical ventilation and how their interpretation may influence the management of ventilated patients. The discussion is limited on waveform eye interpretation of the signals without using any intervention which may interrupt the process of mechanical ventilation. DISCUSSION: Flow, volume, and airway pressure may be used to (a) identify the mode of ventilator assistance, triggering delay, ineffective efforts, and autotriggering, (b) estimate qualitatively patient's respiratory efforts, and (c) recognize delayed and premature opening of exhalation valve. These signals may also serve as a tool for gross estimation of respiratory system mechanics and monitor the effects of disease progression and various therapeutic interventions. CONCLUSIONS: Flow, volume, and airway pressure waveforms are valuable real-time tools in identifying various aspects of patient-ventilator interaction.

Data Display↗

Intensive Care Delirium Screening Checklist: evaluation of a new screening tool.

OBJECTIVE: Delirium in the intensive care unit is poorly defined. Clinical evaluation is difficult in the setting of unstable, often intubated patients. A screening tool may improve the detection of delirium. METHOD: We created a screening checklist of eight items based on DSM criteria and features of delirium: altered level of consciousness, inattention, disorientation, hallucination or delusion, psychomotor agitation or retardation, inappropriate mood or speech, sleep/wake cycle disturbance, and symptom fluctuation. During 3 months, all patients admitted to a busy medical/surgical intensive care unit were evaluated, and the scale score was compared to a psychiatric evaluation. RESULTS: In 93 patients studied, 15 developed delirium. Fourteen (93%) of them had a score of 4 points or more. This score was also present in 15 (19%) of patients without delirium, 14 of whom had a known psychiatric illness, dementia, a structural neurological abnormality or encephalopathy. A ROC analysis was used to determine the sensitivity and specificity of the screening tool. The area under the ROC curve is 0.9017. Predicted sensitivity is 99% and specificity is 64%. CONCLUSION: This study suggests that the Intensive Care Delirium Screening Checklist can easily be applied by a clinician or a nurse in a busy critical care setting to screen all patients even when communication is compromised. The tool can be utilized quickly and helps to identify delirious patients. Earlier diagnosis may lead to earlier intervention and better patient care.

APACHE↗

An osteoporosis screening tool for Chinese men.

Several osteoporosis risk instruments have been proposed to select women for bone densitometry, but no validated instruments are currently available for men. This study aims to address this deficiency by developing and validating a Male Osteoporosis Screening Tool (MOST) for Chinese men. Two thousand ambulatory men, aged 65 and above, were recruited from the general community in Hong Kong, and a cohort of 1,970 men with valid total hip and lumbar spine dual-energy X-ray absorptiometry (DXA) measurements was included in the current analysis. A 60% random sample was selected as the training sample for developing the screening tool, and the remaining 40% constituted the validation sample. Logistic regression and receiver operating characteristic (ROC) analysis were used to identify the simplest combination of risk factors to be included in the screening tool for predicting osteoporosis at the femoral neck, total hip, or lumbar spine. Body weight and quantitative ultrasound index (QUI) were found to contribute significantly to the area under the ROC curve (AUC), yielding an AUC of 0.823 in the training sample. The resulting MOST had a sensitivity of 94% and a specificity of 46% when using a cutoff score of 3. MOST had an AUC of 0.839 in the validation sample. The risk of osteoporosis was 1% among those with MOST scores < or = 2, but 72% among those with MOST scores > 7. Using a cutoff of 3, the negative predictive value was 97.5% which suggests that the 42% with MOST scores < or = 3 may be accurately screened out as being without osteoporosis, thus saving two fifths of our DXA resources. The positive predictive value was 72% when using a cutoff of 7, implying that MOST cannot replace DXA for case-finding purposes. Nevertheless, for resource allocation and patient satisfaction, it is prudent and economical to offer DXA screening first to the 6% with MOST scores > 7.

Aged↗