Specific Measurement Method Research Articles

Development of Enzyme-Linked Immunosorbent Assay System for PEDF and its Clinical Utility

Pigment epithelium-derived factor (PEDF) is reported to play a protective role against diabetic vascular complications through its anti-oxidative properties. However, since a commercially available kit is not suitable for measurement of serum PEDF in humans, kinetics and regulation of serum PEDF are not known in these devastating disorders. Therefore, we developed a simple, specific and reliable method for measurement of serum PEDF in humans using a competitive enzyme-linked immunosorbent assay (ELISA) system. Assay linearity was shown intact with 50~300-fold dilution of urea-pretreated serum by phosphate-buffered saline. The recovery ratio of added recombinant human PEDF in serum was 94.2 +/- 1.7 %. Inter- and intra-assay coefficient of variations of the ELISA were 4.7 and 7.3 %, respectively. When we measured serum PEDF levels in a general population, PEDF levels were elevated in proportion to the accumulation of the number of the components of the metabolic syndrome. Further, the percent changes in serum levels of PEDF during 1-year observational periods were positively correlated with those of body mass index (BMI) in patients with type 2 diabetes. In addition, PEDF mRNA levels in cultured adipocytes were increased in parallel to the BMI values of subjects from which adipocytes were derived, especially in omental adipocytes. These observations suggest that PEDF is generated from adipose tissues and could be increased as a counter-system against vascular cell damage in humans. PEDF may be one of the useful biomarkers for vascular injury in high-risk patients.

Utilization of the Electrostatic Force Microscopy for Detection Filler Grains in Nanocomposites

1. IntroductionDynamic development of new class of the materialsopens the opportunity to introduce breaking-throughtechnologies in the industry. Specific properties of suchmaterialareconnectedtothenanometer-sizeobjectsorfeatures. Theirdevelopmentispossibleduetohighres-olution diagnostic techniques. In many investigations,the application of specific measurement method is vi-talforobtainingspecificinformation. Thereforeonecanstill observe progress in that field [1–3]. One of inter-estingmaterialsistheepoxymatrixwiththesilicafillerof nanometer size (typically 10–30 nm of diameter). Itcanbeusedasanewgenerationdielectricmaterial. Itspermittivity and dissipation factor can be much betterthaninproductspresentonthemarket[4]. Thedevicesmadeofsuchmaterialcanbemoreresistiveforwearandclimatic conditions and require lower energy consump-tionduringproductionprocess. Inordertocorrelatethetechnologyandmacroscopicmeasurementresults[5],oneshouldknowthenanofillerdistribution.2. MaterialdiagnosticproblemsTo provide information about nanoparticles presenceand their distribution in the matrix, high-resolutionimaging methods are desired. The atomic force mi-croscopy (AFM) method proved its efficiency in vari-ousapplicationsincludingdielectricmaterialsinvestiga-tions. Althoughthistechniquegenerallydeliversthein-formationonlyaboutthesurfaceproperties,onecanusethe modes which can detect interaction of few tens ofnanometersrange,thereforeinsightintothebulkcanbeobtained. AsshowninFig.1a,theshort-distanceinter-action cannot reveal presence of the objects, if at least

Serum steroid profiling for Congenital Adrenal Hyperplasia using liquid chromatography–tandem mass spectrometry

Background Diagnosis of Congenital Adrenal Hyperplasia (CAH) is based on the quantification of 17-hydroxyprogesterone (17-OHP), usually by immunoassay. During the neonatal period the specificity of screening for CAH by blood spot 17-OHP immunoassay is low. High false-positive rates result in a relatively high demand for a second-tier serum confirmation test. A robust, specific and selective method for measurement of cortisol, 21-deoxycortisol, 11-deoxycortisol, 4-androstene-3,17-dione (A4) and 17-OHP in serum has been developed. The method involves a simple extraction procedure and a fast analysis using ultra-performance liquid chromatography–tandem mass spectrometry (UPLC/MS/MS). Methods The steroids were extracted from 50 µl of serum using methyl-tert-butyl-ether. Analysis was performed on a UPLC tandem quadrupole mass spectrometer system in positive mode electrospray ionization and multiple reaction monitoring acquisition. Results The assay was linear over each analyte concentration range with all correlation coefficients ( r 2) > 0.996. Inter- and intra-day CVs were ≤ 10% across the analytical range. In addition simultaneous measurement of the full range of steroids on the pathway to cortisol allows confirmation of the affected steroidogenic enzyme. Conclusions A second-tier test for the confirmation of CAH has been developed. The method allows for detection and quantification of 5 steroids related to CAH over the range of the clinical assay with good linearity, sensitivity and precision.

Measurement of plasma renin concentration in cats by use of a fluorescence resonance energy transfer peptide substrate of renin

To evaluate the use of a commercially available 5-carboxyfluorescein-based, intramolecularly quenched, fluorescence resonance energy transfer (FRET) peptide substrate of renin for measurement of plasma renin concentration in cats. Plasma samples obtained during a previous study of renal autograft ischemia-reperfusion injury in 10 cats and samples of fetal bovine serum containing recombinant human renin (rh-renin). Experiments involving samples of fetal bovine serum containing rh-renin were conducted to identify a suitable control vehicle, optimal substrate concentration, and appropriate duration of incubation. With the use of the identified assay conditions, a standard curve was constructed to allow conversion of relative fluorescent units into values of renin concentration (ng/mL). Subsequently, plasma samples obtained from cats before and after renal autograft ischemia-reperfusion injury were assayed to determine endogenous renin concentration. Under conditions of a 1:50 substrate dilution and 4-hour incubation period, the assay detected small amounts of rh-renin in fetal bovine serum. A linear relationship (R(2) = 0.996) between the relative fluorescent units generated and exogenous rh-renin concentration was evident. The assay detected renin in plasma samples obtained from cats after renal autograft ischemia-reperfusion, and renin concentrations on days 1 and 2 after transplant differed significantly. The study data indicated that the assay involving the FRET peptide substrate of renin is potentially a rapid and specific method for measurement of plasma renin concentration in cats.

Sample cleanup-free determination of mycophenolic acid and its glucuronide in serum and plasma using the novel technology of ultra-performance liquid chromatography–electrospray ionization tandem mass spectrometry

Mycophenolic acid (MPA) is an immunosuppressant drug which powerfully inhibits lymphocyte proliferation. Since the early 1990s it has been used to prevent rejection in organ transplantation. The requirement of therapeutic drug monitoring shown in previous studies raises the necessity of acquiring accurate and sensitive methods to measure MPA and its major metabolite mycophenolic acid glucuronide (MPAG). The authors developed a sample cleanup-free, rapid, and highly specific method for simultaneous measurement of MPA and MPAG in human plasma and serum using the novel technology of ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometry. MPA- and MPAG-determinations were performed during a 2.0-min run time. Multiple calibration curves for the analysis of MPA and MPAG exhibited consistent linearity and reproducibility in the range of 0.05-100 (r>0.999) mg L(-1) and 4-4000 mg L(-1) (r>0.999), respectively. Limits of Detection were 0.014 mg L(-1) for MPA and 1.85 mg L(-1) for MPAG. Lower Limits of Quantification were 0.05 mg L(-1) for MPA and 2.30 mg L(-1) for MPAG. Interassay imprecision was <10% for both substances. Mean recovery was 103.6% (range 78.1-129.7%) for MPA and 111.1% (range 73.0-139.6%) for MPAG. Agreement was good for MPA and MPAG between the presented method and a validated HPLC-MS/MS method. The Passing-Bablok regression line for MPA and MPAG was HPLC-MS/MS=1.14 UPLC-MS/MS-0.14 [ mg L(-1)], r=0.96, and HPLC-MS/MS=0.77 UPLC-MS/MS+0.50 [ mg L(-1)], r=0.97, respectively. This sample cleanup-free and robust LC-MS/MS assay facilitates the rapid, accurate and simultaneous determination of MPA and MPAG in human body fluids.

Using intellectual capital management for facilitating organizational change

PurposeThis paper aims to examine the role of intellectual capital (IC) management in an organizational change process. The purpose is to obtain new empirical findings and an enhanced understanding of the role of IC in an organizational change process is obtained.Design/methodology/approachFirst, the existing literature on change management and IC management is reviewed. Then, empirical research is carried out in terms of an action research project on IC development processes in three case companies. These processes and their outcomes are evaluated using interviews.FindingsAn IC model can be a useful tool for change management as it helps to ensure the alignment of the change content with the strategic goals of the organization.Research limitations/implicationsThe study is based on applying the Danish IC management model in Finnish companies. Different results might have been obtained if another IC management model had been used or the companies representing other countries and cultures had been selected.Practical implicationsThe paper demonstrates that IC tools can be useful for promoting organizational change processes. However, more important than the specific conceptual model or measurement method chosen seems to be the process of organizational representatives connecting their contextual and idiosyncratic understandings to the model.Originality/valueThis paper makes a contribution to the existing knowledge by reporting and critically evaluating the implementation of dynamic IC management practices. This is a contribution to the existing conceptually oriented literature on the topic. In addition, the analysis of empirical IC management projects applied in a change context has novelty value.

A new easy method for specific measurement of active myeloperoxidase in human biological fluids and tissue extracts

The SIEFED (“Specific Immunological Extraction Followed by Enzymatic Detection”) method already developed for the specific detection of the activity of equine myeloperoxidase (MPO) was adapted for the specific measurement of active human MPO in biological fluids or tissue extracts. The method consists of the extraction of MPO from aqueous solutions by immobilized anti-MPO antibodies followed by a washing (to eliminate the extraction medium and the biological fluid with their possible interfering molecules) and the measurement of the activity of MPO with a detection system containing a fluorogenic substrate, H 2O 2 and nitrite ions as reaction enhancer. The SIEFED was applied to study active MPO in human biological fluids (plasma, bronchoalveolar lavage fluid and supernatant from carotids extracts). The SIEFED for human MPO has a sensitivity limit of 0.080 mU/mL and showed good precision with intra- and inter-assay coefficients of variation below 10 and 20% respectively within a broad range of MPO activities establish from 0.156 to 473 mU/mL. The SIEFED for human MPO will be useful for the specific detection of active MPO in complex fluids and can be complementary to an ELISA to determine an active/total MPO ratio in healthy volunteers and patients especially in case of chronic or acute inflammatory diseases.

The Multitrait-Multimethod Matrix at 50!

Fifty years ago, in 1959, Campbell and Fiske published one of the most influential papers in psychology. In their article Convergent and discriminant validation by the multitraitmultimethod matrix, they argued that it is not sufficient to consider one single operationalization of one construct for purposes of test validation but that multiple measures of multiple constructs are necessary. Campbell and Fiske recommended using at least two methods that are as different as possible for measuring the constructs. Moreover, Campbell and Fiske made clear that it is not possible to get a measure of a trait that is free of method-specific influences. Whenever, in science, we measure a construct (a trait) we have to use a specific measurement method. Therefore, it is the trait and the method that influence the observed score simultaneously. In order to separate methodfrom traitspecific influences, it is thus always necessary to consider more than one trait and more than one method in the validation process. Campbell and Fiske proposed the multitraitmultimethod (MTMM) matrix for analyzing the convergent and discriminant validity. The MTMM matrix consists of the correlations between all multiple measures representing the different traits measured by the different methods. These correlations can be evaluated by several criteria that have been developed by Campbell and Fiske. If the different measures of the same construct are highly correlated, this proves convergent validity. If the different measures of one construct are not correlated with the measures of another construct, this indicates discriminant validity. Campbell and Fiske’s article had and has an enormous influence on psychology (Eid & Diener, 2006). It is the most often cited paper that has ever been published in Psychological Bulletin (Sternberg, 1992). To date, it has been cited 4,735 times (Social Science Citation Index, February 27, 2009, 3:41 pm), and its citation rate is increasing. Their article does not only have an important impact on test validation studies but also has a strong impact on methodological research as many researchers have developed new approaches for analyzing MTMM data and tried to overcome some of the problems and limitations that are related to former approaches of analyzing MTMM matrices. This special issue is dedicated to honoring Campbell and Fiske’s influential work. It presents three different modern approaches for analyzing MTMM data. All contributors use the same data set illustrating their approaches. This enables readers to concentrate on the comparison of the different approaches with respect to the way convergent and discriminant validity can be analyzed as well as how traitand method-specific influences can be identified and quantified. The data consists of three personality traits (extraversion, neuroticism, and conscientiousness) assessed by three raters (one selfand two peer raters). Each scale consists of four items (adjectives such as talkative, conscientious, etc.) that were rated on a five-point scale. The sample size is n = 481. Maas, Lensvelt-Mulders, and Hox show in their contribution A multilevel multitrait-multimethod analysis how multilevel models can be applied to analyze MTMM data. Oort presents an application of Three-mode models for multitrait-multimethod data, and Nussbeck, Eid, Geiser, Courvoisier, and Lischetzke show how the data can be analyzed by A CTC(M-1) model for different types of raters. Finally, Hofling, Schermelleh-Engel, and Mossbrugger compare these approaches in their contribution Analysing multitrait-multimethod (MTMM) data: A comparison of three approaches. We hope that the readers will enjoy seeing how this important field of methodological research has developed over the last years.

Tracking Objects in Depth using Size Change

Event Abstract Back to Event Tracking Objects in Depth using Size Change Chen Zhang1* and Julian Eggert2 1 Darmstadt University of Technology, Control Theory and Robotics Lab, Germany 2 Honda Research Institute Europe GmbH, Germany Tracking an object in depth is an important task, since the distance to an object often correlates with an imminent danger, e.g. in the case of an approaching vehicle. A common way to estimate the depth of a tracked object is to utilize binocular methods like stereo disparity. In practice, however, depth measurement using binocular methods is technically expensive due to the need of camera calibration and rectification. In addition, higher depths are difficult to estimate because of the inverse relationship between disparity and depth.Here, we introduce an alternative approach for depth estimation, Depth-from-Size. This is a human-inspired monocular method where the depth is gained by utilizing the fact that object depth is proportional to the ratio of object physical size and object retinal size. Since both the physical size and the retinal size are unknown terms, they have to be measured and estimated together with the depth in a mutually interdependent manner. For each of the three terms specific measurement and estimation methods are probabilistically combined. This results in probability density functions (pdfs) at the output of three components for measuring and estimating these three terms, respectively.In every processing step, we use a 2D tracking system for first obtaining the object’s 2D position in the current monocular 2D image. On the position where the target object is found, the scaling factor of the object retinal size is measured by a pyramidal Lucas-Kanade-algorithm. In our setting, the object retinal size is the only observable subject to frequent measurements, whereas physical size and depth are internal states that have to be inferred by the system according to the constraint - depth / focal length = physical size / retinal size - that couples the three terms. Bayesian estimators are used to estimate the pdfs of the retinal size and the depth, whereas the physical size is gained by a mean estimator, since it is assumed to remain constant over time. Additional measurement inputs for the physical size and the depth are optional, acting as correcting evidences for these both terms.Measuring only the retinal size leaves us with an inherent ambiguity in the system, so that either the physical size or the depth must become available once at initialization. In our system, for this purpose we used a known object size or depth information gained by other depth cues like stereo disparity.The performance of the proposed approach was evaluated in two scenarios: An artificial with ground truth and a real-world scenario. In the latter, depth estimation performance of this system is compared with that of a directly measured stereo disparity. The evaluation results show that this approach is a reliable alternative to the standard stereo disparity approach for depth estimation with several advantages: 1) simultaneous estimation of depth, physical size and retinal size; 2) no stereo camera calibration and rectification; 3) good depth estimation at higher depth ranges for large objects. Conference: Bernstein Conference on Computational Neuroscience, Frankfurt am Main, Germany, 30 Sep - 2 Oct, 2009. Presentation Type: Poster Presentation Topic: Abstracts Citation: Zhang C and Eggert J (2009). Tracking Objects in Depth using Size Change. Front. Comput. Neurosci. Conference Abstract: Bernstein Conference on Computational Neuroscience. doi: 10.3389/conf.neuro.10.2009.14.026 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 25 Aug 2009; Published Online: 25 Aug 2009. * Correspondence: Chen Zhang, Darmstadt University of Technology, Control Theory and Robotics Lab, Darmstadt, Germany, czhang@rtr.tu-darmstadt.de Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Chen Zhang Julian Eggert Google Chen Zhang Julian Eggert Google Scholar Chen Zhang Julian Eggert PubMed Chen Zhang Julian Eggert Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Accuracy of biometry in pediatric cataract extraction with primary intraocular lens implantation

To determine the accuracy of predicted postoperative refractive outcomes in pediatric patients having cataract surgery with intraocular lens (IOL) implantation and to compare them with other variables historically considered important in cataract surgery. Tertiary care referral hospital. This retrospective review comprised 203 eyes of 153 consecutive pediatric patients (< or = 18 years old) having cataract extraction with primary posterior chamber IOL implantation in the capsular bag. All cases were performed by 1 of 2 surgeons, and all refractions were performed manually by an experienced pediatric ophthalmologist using a retinoscope. In all patients, the mean absolute value (MAE) of the prediction error was 1.08 diopters (D) +/- 0.93 (SD). Age at time of surgery and corneal (K) mean curvature were significantly correlated with the absolute value of the prediction error (P = .0006 and P = .0088, respectively). A multiple regression model showed that age at time of surgery and K mean curvature were the only 2 variables significantly associated with MAE; axial length, formula, surgeon, and A-scan type were not significantly associated with prediction error. Data from 203 consecutive primary pediatric IOL implantations showed the heterogeneous nature of the variables involved in predictions of refractive outcomes in this population. The complexities of this issue support the need for specific methods of measurement and an IOL calculation formula for the pediatric population.

Temporal suppression and augmentation of click-evoked otoacoustic emissions

This study investigates temporal suppression of click-evoked otoacoustic emissions (CEOAEs), occurring when a suppressor-click is presented close in time to a test-click (e.g. 0–8 ms). Various temporal suppression methods for examining temporal changes in cochlear compression were evaluated and measured here for seven subjects, both for short- and long-latency CEOAEs. Long-latency CEOAEs (duration >20 ms) typically indicate the presence of synchronised spontaneous otoacoustic emissions (SSOAEs). Temporal suppression can only be linked to changes in CEOAE-compression if the suppressor-click affects the CEOAE magnitude. Phase changes induced by the suppressor-click were shown to bias suppression in two ways: (i) when a specific asymmetric measurement method was used and (ii) when synchronisation between the CEOAE and the click-stimuli was incomplete. When such biases were eliminated, temporal suppression and augmentation (the opposite effect) were observed and shown to be subject-dependent. This indicates that the nonlinearity underlying temporal suppression can work in a more (i.e., suppressed) or less (i.e., augmented) compressive state, depending on the inter-click interval and the subject under test. Temporal suppression was shown to be comparable for CEOAEs and SSOAEs, indicating similar underlying cochlear nonlinear mechanisms. This study contributes to a better understanding of the temporal properties of cochlear dynamics.

Measurement of tacrolimus in blood with the method of high performance liquid chromatography-mass spectrometry

To develop a simple, precise, and specific method for measurement of tacrolimus in whole blood. Tacrolimus was extracted from the blood samples of 40 patients treated with tacrolimus by liquid-liquid extraction. Then high performance liquid chromatography-mass spectrometry (HPLC/MS) was used, with ascomycin as internal marker, to isolate the tacrolimus and measure the concentration thereof. The standard curve was drawn. The linearity of quantitative measurement was determined. Detection limits test, CV test, and recovery test were performed. ELISA was used simultaneously. The results of these 2 methods were compared. A good standard curve was drawn based on the results of HPLC/MS assay (y = 0.2464x + 0.0082, R2 = 0.9996). The detected data were highly related to the detected concentrations. The linearity range was 0.100 - 40.000 ng/ml (y = 1.0294x-0.035, R2 = 0.9998). The detection limit of the assay was 0.100 ng/ml. The CV values by this assay were basically < 5%. The recovery rate ranged 95.67% - 98.30% for tacrolimus over the range 0.300 - 16.016 ng/ml. There was a linear correlation (y = 1.0172x + 0.3742, R2 = 0.9630) between the assay results by HPLC/MS to ELISA in blood. The measurement value of ELISA was (19 +/- 9), significantly higher than that of HPLC/MS (18 +/- 9, P < 0.01). This newly developed method of HPLC/MS is simple, precise, and specific and with lower cost, it can be used in the clinical practice and experimental study on tacrolimus.

Procurement process maturity: key to performance measurement

Performance measurement has been receiving increased attention in public organizations. As performance measurement systems begin to take on a central focus by public sector organizations, the challenges of measuring and improving critical organizational processes continue to increase in importance. Furthermore, as the procurement process continues to gain critical importance in public sector organizations, the need to apply specific performance measurement methods to measure and improve the procurement processes is essential for mission success. This article introduces the Contract Management Maturity Model (CMMM) as a method for assessing, measuring, and improving an organization’s procurement processes. The results of the case study describe the organizational benefits of using a contract management maturity model as a performance measurement, as well as a process improvement method.

Method for specific heat measurement of thick film layers

A method of simple implementation is presented for the measurement of the specific heat of thick film layers. It has to do with a system, operating at room temperature in accordance with a dynamic non-adiabatic technique, whose function regards the transient phase of a heating process between two steady states. It consists of a compact ensemble of three elements: a small plate thermistor serving as a heater, an alumina disk as a sample support and a tiny calibrated thermistor as a temperature sensor. The voltage across the heater and temperature from the sensor are monitored. An accurate evaluation of the dissipated energy allows the absorbed energy, responsible for the temperature variation, to be brought out. That makes possible the determination of thermal capacity of a small mass material deposited on a substrate. The achievement of the result simply involves the recording of a voltage and temperature during the transient phase, a fitting procedure with given analytical functions and a mathematical elaboration. The measuring system, its calibration procedure together with the transient operation are described. An application concerning screen-printed ferroelectric thick films on alumina substrate is presented and the results discussed.

Effect of Formalin Fixation on Tumor Size Determination in Stage I Non-Small Cell Lung Cancer

Tumor size is an important prognostic factor in non-small cell lung cancer (NSCLC), but the American Joint Committee on Cancer staging system does not mandate a specific measurement method. Moreover, measuring fresh specimens and formalin-fixed specimens may yield disparate results. Our goal was to evaluate this disparity for stage I NSCLC. We enrolled 401 patients with stage I NSCLC who underwent surgical interventions and follow-up in our hospital between 1993 and 2002. Tumors invading visceral pleura, involving the main bronchus, or associated with atelectasis or obstructive pneumonitis were excluded. Tumor size was measured immediately after resection by surgeons and after formalin fixation by pathologists. Patients were assigned to one of three groups. Group 1 included 201 patients with tumors of 3 cm or less as indicated by both operation notes and pathology reports. Group 2 included 160 patients with tumors larger than 3 cm by both records. Group 3 included 40 patients with tumors larger than 3 cm according to operation notes but 3 cm or less according to pathology reports. Survival rates were compared. Mean follow-up was 58 months. Five-year survival was 70.1% in group 1, 49.1% in group 2, and 51.1% in group 3. As expected, there was a significant survival difference between groups 1 and 2 (p < 0.001); however, there was also a difference between groups 1 and 3 (p = 0.006). Formalin fixation may cause tumor shrinkage and migration from T2 to T1. For accurate tumor staging, size measurements should be performed immediately after resection instead of after formalin fixation. TNM staging should specify how to measure tumor size and the specimen status to be measured.

Quantification of the rat spinal microglial response to peripheral nerve injury as revealed by immunohistochemical image analysis and flow cytometry

Microgliosis is implicated in the pathophysiology of several neurological disorders, including neuropathic pain. Consequently, perturbation of microgliosis is a mechanistic and drug development target in neuropathic pain, which highlights the requirement for specific, sensitive and reproducible methods of microgliosis measurement. In this study, we used the spinal microgliosis associated with L5 spinal nerve transection and minocycline-induced attenuation thereof to: (1) evaluate novel software based semi-quantitative image analysis paradigms for the assessment of immunohistochemical images. Microgliosis was revealed by immunoreactivity to OX42. Several image analysis paradigms were assessed and compared to a previously validated subjective categorical rating scale. This comparison revealed that grey scale measurement of the proportion of a defined area of spinal cord occupied by OX42 immunoreactive cells is a robust image analysis paradigm. (2) Develop and validate a flow cytometric approach for quantification of spinal microgliosis. The flow cytometric technique reliably quantified microgliosis in spinal cord cell suspensions, using OX42 and ED9 immunoreactivity to identify microglia.The results suggest that image analysis of immunohistochemical revelation of microgliosis reliably detects the spinal microgliosis in response to peripheral nerve injury and pharmacological attenuation thereof. In addition, flow cytometry provides an alternative approach for quantitative analysis of spinal microgliosis elicited by nerve injury.

Psychosomatic obstetrics and gynecology – a neglected field?

The purpose of the review was to evaluate recent studies in the field of psychosomatic obstetrics and gynecology for new directions in research. Psychosomatic obstetrics and gynecology covers a wide canvas, and the present review shows extensive research in some areas and large gaps in others. There have been significant developments in the field of pregnancy-related psychiatric morbidity, particularly anxiety states and posttraumatic stress disorder; however, there is a need for better and more specific measurement and screening methods. Among gynecological conditions there have been advances in research on chronic pelvic pain and gynecological cancers, but most other gynecological conditions have been neglected. What is encouraging is the increasing amount of research from the developing world and the focus on common reproductive conditions based on community studies. In addition, in light of the recent findings and concerns related to hormone replacement therapy, research on mood disorders related to menopause is increasing, with refinement in measurements, definitions, and treatment methods. While studies on prevalence and risk factors abound, what is missing are well conducted intervention studies. The next phase of research should focus on preventive and intervention studies that are easily adaptable the world over.

A Specific Method for Measurement of Equine Active Myeloperoxidase in Biological Samples and in in Vitro Tests

An original method called SIEFED (specific immunological extraction followed by enzymatic detection) was developed for the specific detection of the activity of equine myeloperoxidase (MPO). The method consists of the extraction of MPO from aqueous solutions by immobilized anti-MPO antibodies followed by washing (to eliminate proteins and interfering molecules) and measurement of MPO activity using a detection system containing a fluorogenic substrate, hydrogen peroxide, and nitrite as reaction enhancer. The SIEFED technique was applied to study active MPO in horse biological fluids and the effects of 2 polyphenolic molecules, curcumin and resveratrol, on MPO activity. The detection limit of the SIEFED was 0.23 mU/ml. The SIEFED exhibited good precision with intra-assay and interassay coefficients of variation below 10% and 20%, respectively, for MPO activities ranging from 0.25 to 6.4 mU/ml. The activity of MPO was generally higher than 1 mU/ml in the fluids collected from horses with inflammatory diseases. Curcumin and resveratrol exerted a dose-dependent inhibition on MPO activity and, as they were removed before the enzymatic detection of MPO, the results suggest a direct drug-nzyme interaction or an enzyme structure modification by the drug. The SIEFED is a new tool that would be useful for specific detection of active MPO in complex media and for selection of MPO activity modulators.

The Development of the Toronto Structured Interview for Alexithymia: Item Selection, Factor Structure, Reliability and Concurrent Validity

Background: Assessments of personality constructs increasingly use self-report and structured interview instruments, which allow for a multimethod measurement approach and decrease specific measurement method bias. The aim of this study was to develop a valid and reliable structured interview for assessing the alexithymia construct. Methods: Sixty interview questions were written initially, each with a set of scoring criteria and prompts and probes to elicit information assisting in the scoring of the respondents’ answers. Results: After pilot testing, the number of questions was reduced to 43, which were administered to 136 community participants and 97 psychiatric outpatients. A series of item and scale analyses further reduced the item pool to 24 items. Principal component analysis and confirmatory factor analysis of these 24 items revealed preliminary evidence of a hierarchical, four-factor structure, with four lower factors nested within two higher-order latent factors. This structural configuration resulted in the Toronto Structured Interview for Alexithymia (TSIA) with two domain scales and four facet scales. The TSIA and its six scales demonstrated acceptable levels of interrater, internal, and retest reliability. The TSIA and its scales correlated modestly but significantly with the 20-item Toronto Alexithymia Scale and its three factor scales, providing some support for the concurrent validity of this interview. Conclusion: The TSIA appears to be a promising structured interview for assessing alexithymia.

Nitric oxide mediated oxidative stress injury in rat skeletal muscle subjected to ischemia/reperfusion as evaluated by chemiluminescence

The involvement of nitric oxide (*NO) in oxidative stress in the rat gastrocnemius muscle subjected to ischemia/reperfusion injury was investigated using a specific and sensitive chemiluminescence (CL) method for measurement of both membrane lipid peroxide and total tissue antioxidant capacity (TRAP). In addition, inhibitors of nitric oxide synthase enzymes were used. The CL time-course curve increased dramatically after 1, 2, and 4 h of reperfusion, reaching values about 12 times higher than those of both control and ischemic rats. Initial velocity (V0) increased from 13.6 cpm mg protein(-1) min(-1) in the ischemic group, to 7341-8524 cpm mg protein(-1) min(-1) following reperfusion. The administration of L-NAME prior to reperfusion significantly reduced (p<0.007) the time-course of the CL curve, decreasing the V(0) value by 51% and preventing antioxidant consumption for 1h following reperfusion. No significant change in CL time-course curve and TRAP values were observed with aminoguanidine treatment. On contrary, after 4h following reperfusion, pre treatment with aminoguanidine led to a significant decrease (p < 0.0001) in the time-course of the CL curve, where V0 decreased by 75% and TRAP returned to control levels. No significant change in CL time-course curve and TRAP values were observed with L-NAME treatment. When RT-PCR was carried out with an iNOS-specific primer, a single band was detected in RNA extracted from muscle tissue of only the 4 h ischemia/4 h reperfusion group. No bands were found in either the control, 4 h ischemia or 4 h ischemia/1 h reperfusion groups. Based on these results, we conclude that *NO plays an important role in oxidative stress injury, possibly via -ONOO, in skeletal muscle subjected to ischemia/reperfusion. Our results also show that cNOS isoenzymes are preferentially involved in *NO generation at the beginning of reperfusion and that iNOS isoenzyme plays an important role in reperfusion injury producing *NO later in the process.