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Results for “Repeatability and Reproducibility”

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PubMed1990

Graphical analysis of reversible radioligand binding from time-activity measurements applied to [N-11C-methyl]-(-)-cocaine PET studies in human subjects.

A graphical method of analysis applicable to ligands that bind reversibly to receptors or enzymes requiring the simultaneous measurement of plasma and tissue radioactivities for multiple times after the injection of a radiolabeled tracer is presented. It is shown that there is a time t after which a plot of integral of t0ROI(t')dt'/ROI(t) versus integral of t0Cp(t')dt'/ROI(t) (where ROI and Cp are functions of time describing the variation of tissue radioactivity and plasma radioactivity, respectively) is linear with a slope that corresponds to the steady-state space of the ligand plus the plasma volume,.Vp. For a two-compartment model, the slope is given by lambda + Vp, where lambda is the partition coefficient and the intercept is -1/[kappa 2(1 + Vp/lambda)]. For a three-compartment model, the slope is lambda(1 + Bmax/Kd) + Vp and the intercept is -[1 + Bmax/Kd)/k2 + [koff(1 + Kd/Bmax)]-1) [1 + Vp/lambda(1 + Bmax/Kd)]-1 (where Bmax represents the concentration of ligand binding sites and Kd the equilibrium dissociation constant of the ligand-binding site complex, koff (k4) the ligand-binding site dissociation constant, and k2 is the transfer constant from tissue to plasma). This graphical method provides the ratio Bmax/Kd from the slope for comparison with in vitro measures of the same parameter. It also provides an easy, rapid method for comparison of the reproducibility of repeated measures in a single subject, for longitudinal or drug intervention protocols, or for comparing experimental results between subjects. Although the linearity of this plot holds when ROI/Cp is constant, it can be shown that, for many systems, linearity is effectively reached some time before this. This analysis has been applied to data from [N-methyl-11C]-(-)-cocaine ([11C]cocaine) studies in normal human volunteers and the results are compared to the standard nonlinear least-squares analysis. The calculated value of Bmax/Kd for the high-affinity binding site for cocaine is 0.62 +/- 0.20, in agreement with literature values.

Brain↗

PubMed2000

Quantitation of minimal residual disease in acute promyelocytic leukemia patients with t(15;17) translocation using real-time RT-PCR.

We took advantage of a recently developed system allowing performance of real-time quantitation of polymerase chain reaction to develop a quantitative method of measurement of PML-RARalpha transcripts which are hallmarks of acute promyelocytic leukemia (APL) with t(15;17) translocation. Indeed, although quantitation of minimal residual disease has proved to be useful in predicting clinical outcome in other leukemias such as chronic myeloid leukemia or acute lymphoblastic leukemia, no quantitative data have been provided in the case of APL. We present here a method for quantitation of the most frequent subtypes of t(15;17) transcripts (namely bcr1 and bcr3). One specific forward primer is used for each subtype in order to keep amplicon length under 200 bp. The expression of PML-RARalpha transcripts is normalized using the housekeeping porphobilinogen deaminase (PBGD) gene. This technique allows detection of 10 copies of PML-RARalpha or PBGD plasmids, and quantitation was efficient up to 100 copies. One t(15;17)-positive NB4 cell could be detected among 106 HL60 cells, although quantitation was efficient up to one cell among 105. Repeatability and reproducibility of the method were satisfying as intra- and inter-assay variation coefficients were not higher than 15%. The efficiency of the method was finally tested in patient samples, showing a decrease of the PML-RARalpha copy number during therapy, and an increase at the time of relapse.

Chromosomes, Human, Pair 15↗

PubMed2003

False leukemia-lymphoma cell lines: an update on over 500 cell lines.

Human leukemia-lymphoma (LL) cell lines represent an extremely important resource for research in a variety of fields and disciplines. As the cell lines are used as in vitro model systems in lieu of primary cell material, it is crucial that the cells in the culture flasks faithfully correspond to the purported objects of study. Obviously, proper authentication of cell line derivation and precise characterization are indispensable requirements to use as model systems. A number of studies has shown an unacceptable level of LL cell lines to be false. We present here the results of authenticating a comprehensively large sample (n = 550) of LL cell lines mainly by DNA fingerprinting and cytogenetic evaluation. Surprisingly, near-identical incidences (ca 15%) of false cell lines were observed among cell lines obtained directly from original investigators (59/395: 14.9%) and from secondary sources (23/155: 14.8%) implying that most cross-contamination is perpetrated by originators, presumably during establishment. By comparing our data with those published, we were further able to subclassify the false cell lines as (1) virtual: cross-contaminated with and unretrievably overgrown by other cell lines during initiation, never enjoying independent existence; (2) misidentified: cross-contaminated subsequent to establishment so that an original prototype may still exist; or (3) misclassified: unwittingly established from an unintended (often normal) cell type. Prolific classic leukemia cell lines were found to account for the majority of cross-contaminations, eg CCRF-CEM, HL-60, JURKAT, K-562 and U-937. We discuss the impact of cross-contaminations on scientific research, the reluctance of scientists to address the problem, and consider possible solutions. These findings provide a rationale for mandating the procurement of reputably sourced LL cell lines and their regular authentication thereafter.

Cell Lineage↗

PubMed1998

Alpha 1-acid glycoprotein affinity columns for the clean-up of adrenergic drugs.

alpha 1-Acid glycoproteins (AAGs) have a structure resembling beta-adrenergic receptors and bind several basic drugs in plasma. Chromatographic columns were prepared by linking epsilon-NH2 groups of AAG lysines to a Sepharose 4B support, in order to purify by affinity chromatography adrenergic drugs of possible use in animal production. Loading capacities, binding efficiency, memory effects and matrix interferences from urine samples were studied. The method developed involves sample application in buffered media (pH 7.4), washing with 5 ml of PBS, and elution with 4 ml of 1% v/v acetic acid. Under these conditions no memory effect was observed. Loading capacity is correlated with the physiological plasma binding rate (PB) of the drug. For clenbuterol (PB 50%) and anilino-like related drugs, 5 mg of AAG were able to bind about 15 x 10(-6) g of drug, with a 100% recovery from the column. Repeatability and reproducibility, expressed as RSD, were 4.2 and 5.4%, respectively. The calculated AAG: drug molar ratio was 4.5:1, indicating 22% of the AAG bound to the column retained drug affinity. Among phenolic-like agonists, salbutamol (PB 5%), fenoterol and isoxsuprine hardly interacted, whereas nylidrin, ritodrine and bamethan showed more effective binding. We also checked binding of other drugs of possible use in veterinary medicine. Application of the AAG column to spiked bovine urine revealed a mean recovery of 97.8%; no matrix interferences were observed.

Adrenergic beta-Agonists↗

PubMed1999

Cement for stabilisation of industrial residues containing heavy metals.

A method aimed at decreasing the toxicity of heavy metals [namely, Zn(II) and Cr(III)] in real polluted residues by immobilisation has been developed. The residues were processed either with two cement-type stabilisers or lime. The cement-type stabilisers were Portland cement and Depocrete SM/2 at the self-generated pH (ca. 11) which afforded physical as well as chemical potential for the immobilisation of heavy metals. The other stabiliser, lime, reduced organic compounds, thus favouring the decrease of the chemical oxygen demand (COD) and endowing the residue with better mechanical properties for transport. After leaching the stabilised residues using the standard leaching test [Order 13/10/89, Boletín Oficial del Estado (BOE) 270 10/11/89], three ways for establishing the toxicity of the treated residues were used, namely: (1) the ecotoxicity test using Photobacterium phosphoreum (DIN 38 412); (2) determination of the concentration of heavy metals by atomic absorption spectrometry (AAS); (3) determination of the COD or oxygen required for complete chemical oxidation of a water sample. Portland cement (20%) blended with Depocrete SM/2 (3%) acted as an effective stabiliser for residues containing heavy metals as it increased the ecotoxicity index (EC50) by more than five times. Thus the heavy metal concentration in the leaching liquid was lowered to less than 0.1 mg l-1. The addition of 5% of lime afforded a residue easily transportable from the place of treatment to the landfill. The precision of the method was studied in terms of both repeatability and reproducibility. The values found with respect to EC50 and expressed as the relative standard deviation (RSD) were 1.6% and 5.1%, respectively.

Environmental Pollution↗

PubMed1999

GC-MS determination of steroids related to androgen biosynthesis in human hair with pentafluorophenyldimethylsilyl-trimethylsilyl derivatisation.

An efficient method for the simultaneous determination of eight steroids, androstenedione, dihydrotestosterone (DHT), dehydroepiandrosterone (DHEA), testosterone, androsterone, etiocholanolone, progesterone and pregnenolone, in human hair by gas chromatography-mass spectrometry (GC-MS) using d3-testosterone as internal standard is described. The method involves alkaline digestion, liquid-liquid extraction and subsequent conversion to mixed pentafluorophenyldimethylsilyl-trimethylsilyl (flophemesyl-TMS) derivatives for sensitive analysis in the selected ion monitoring (SIM) mode. This method showed good overall repeatability and reproducibility of 4.88-11.24 and 3.19-9.58%, respectively. For the first time, the quantification of DHT, DHEA and pregnenolone in human hair has been achieved by GC-MS, testosterone was also quantified. The detection of four steroids in hair samples was possible in the concentration range 0.12-8.45 ng g-1. The other four steroids, androstenedione, androsterone, etiocholanolone and progesterone, were not detected. The detection limits for SIM of the steroids varied in the range 0.02-0.5 ng g-1, and the SIM responses were linear with correlation coefficients varying from 0.991 to 0.996 for most of the steroids studied. The concentrations of the four steroids detected were different in male and female hair samples.

Androgens↗

PubMed2000

In situ laser-induced fluorescence (LIF) analysis of petroleum product-contaminated soil samples.

General considerations of the calibrations of in situ measurements are presented and the concept of using an "average oil" with average analysability for calibration purposes is introduced. The in situ analysis of 30 petroleum product-contaminated soil samples with laser-induced fluorescence (LIF) spectroscopy was performed. Compared to an uncontaminated laboratory reference (LR) soil, 23 soil samples exhibited significantly higher LIF signals, so that these soil samples were classified as contaminated. The repeatability and reproducibility of the in situ LIF analysis were investigated. For the calibration of the LIF data, two LR oils (a fuel oil and a crude oil) were employed. The degree of soil contamination with petroleum products ranged from the limit of detection (LOD) for LIF analysis (ca. 100 ppm), or below, to more than 10,000 ppm. The petroleum product concentrations determined with in situ LIF analysis reveal a reasonable correlation with the results of standard IR analysis after extraction of the contaminated soils.

Calibration↗

PubMed1991

Spectrophotometric flow injection determination of trace amounts of thiocyanate based on its reaction with 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol and dichromate: assay of the thiocyanate level in saliva from smokers and non-smokers.

A simple and very sensitive spectrophotometric flow injection (FI) procedure for the determination of trace amounts of thiocyanate is described. The proposed method is based on the reaction between thiocyanate and 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol, which, in 2 mol dm-3 acidic media in the presence of a strong oxidizing agent, produces an intensely coloured product. Several oxidants are potentially applicable, but it is shown that dichromate is preferable. As the reaction product formed is unstable and the signal inherently is recorded on a high background level, it is demonstrated that FI constitutes an ideal method in order to monitor reproducibly and repeatedly the kinetically transient signal that is obtained. Based on optimization by a factorial experimental design, the detection limit of the procedure was found to be 3.5 mumol dm-3, and the standard deviation between samples was 0.16 mumol dm-3. No significant interferences were observed; a 1000-fold excess of cyanide could readily be tolerated within the experimental error. With a sample volume of 50 microliters being injected, the sampling frequency was 60 samples h-1. The system was tested with saliva samples from non-smokers and smokers, and the results show that it is possible to distinguish between these two categories of individuals. As an added benefit, the detection limit of the analytical procedure allows the samples to be diluted 100-fold, so that centrifugation for 5 min is the only preliminary sample preparation that is necessary.

Azo Compounds↗

PubMed2000

Estimation of trace amounts of benzene in solvent-extracted vegetable oils and oil seed cakes.

A new method is presented for the qualitative and quantitative estimation of trace amounts (up to 0.15 ppm) of benzene in crude as well as refined vegetable oils obtained by extraction with food grade hexane (FGH), and in the oil seed cakes left after extraction. The method involves the selection of two solvents; cyclohexanol, for thinning of viscous vegetable oil, and heptane, for azeotroping out trace benzene as a concentrate from the resulting mixture. Benzene is then estimated in the resulting azeotrope either by UV spectroscopy or by GC-MS subject to availability and cost effectiveness of the latter. Repeatability and reproducibility of the method is within 1-3% error. This method is suitable for estimating benzene in vegetable oils and oil seed cakes.

Animal Feed↗

PubMed2000

Splitless large-volume GC-MS injection for the analysis of organophosphorus and organochlorine pesticides in vegetables using a miniaturised ethyl acetate extraction.

A simple, rapid and sensitive multiresidue method has been developed for the determination of ten organophosphorus and organochlorine pesticides, commonly used in crop protection. The analysis uses a miniaturised extraction with ethyl acetate followed by large volume injection (10 microL) GC-EI-MS analysis in SIM (selective ion monitoring) mode. Sensitivity and selectivity of the method were acceptable with limits of detection (LODs) lower than 0.01 mg kg-1, except for endosulfan alpha and beta (0.05 mg kg-1). Average recoveries of between 63-99% were obtained and good linearity was observed in the range from 0.01 to 1.00 mg kg-1. Repeatability and reproducibility studies yielded relative standard deviations lower than 20% in all the cases. The method was applied to the analysis of 110 tomato, pepper and cucumber samples, as part of the monitoring programme of the Association of Producers and Exporters of Fruits and Vegetables of Almería.

Acetates↗

PubMed2001

Gas chromatographic determination of pesticides in vegetable samples by sequential positive and negative chemical ionization and tandem mass spectrometric fragmentation using an ion trap analyser.

A selective and sensitive chromatographic method is described for the determination of nine organochlorine and organophosphorus pesticides in vegetable samples by gas chromatography-mass spectrometry. The proposed method combines the use of positive and negative chemical ionisation and tandem mass spectrometric fragmentation, resulting in a significant increase in selectivity and allowing the simultaneous confirmation and quantification of trace levels of pesticides in complex vegetable matrices. Parameters relative to ionisation and fragmentation processes were optimised to obtain maximum sensitivity. Repeatability and reproducibility studies yielded relative standard deviations lower than 25% in all cases. Identification criteria, such as retention time and relative abundance of characteristic product ions, were also evaluated in order to guarantee the correct identification of the target compounds. The method was applied to real vegetable samples to demonstrate its use in routine analysis.

Gas Chromatography-Mass Spectrometry↗

PubMed2001

Chromatographic analysis of phenethylamine-antihistamine combinations using C8, C18 or cyano columns and micellar sodium dodecyl sulfate-pentanol mixtures.

The chromatographic behaviour of binary and ternary mixtures of several phenethylamines (phenylephrine, phenylpropanolamine, ephedrine, pseudoephedrine and methoxyphenamine) and antihistamines (pheniramine, carbinoxamine, doxylamine, chlorpheniramine, dexchlorpheniramine, dexbrompheniramine, diphenhydramine, tripolidine, azatadine and phenyltoloxamine), found in cough-cold pharmaceutical preparations, was studied using C8, C18 and cyano columns, micellar mobile phases of sodium dodecyl sulfate (SDS) and pentanol and UV detection. Using a C8 column and mobile phases of 0.05 mol l-1 SDS-6% v/v pentanol or 0.15 mol l-1 SDS-2% v/v pentanol at pH 7, more than 30 different phenethylamine-antihistamine combinations can be resolved in < 15 min. Intra- and inter-day repeatabilities and reproducibilities evaluated at three different drug concentrations (0.5, 5 and 25 micrograms ml-1, n = 10) were below 1.6, 2.5 and 2.4%, respectively. The drug amounts found in 18 formulations agreed with those declared by the manufacturers within the tolerance limits, and with those obtained using a mobile phase of 55% v/v methanol at pH 7. No interference was observed from other accompanying drugs such as acetylsalicylic acid, ascorbic acid, betamethasone, bromhexine, caffeine, codeine, dextromethorphan, paracetamol, prednisolone, salicylamide and tartrazine. The proposed procedure has the advantage over the conventional aqueous-organic procedure of using a small amount of organic solvent, which is highly retained in the SDS solution. The efficiencies are also greater. On the other hand, in the micellar system, the retentions of phenethylamines and antihistamines are similar, although the compounds can be easily resolved. In contrast, using the methanol-water mobile phase, the phenethylamines are weakly retained, whereas the antihistamines usually show a high retention.

Chromatography↗

PubMed2001

Automated on-line in-tube solid-phase microextraction followed by liquid chromatography/electrospray ionization-mass spectrometry for the determination of chlorinated phenoxy acid herbicides in environmental waters.

A method for the determination of six chlorinated phenoxy acid herbicides in river water was developed using in-tube solid-phase microextraction (SPME) followed by liquid chromatography/electrospray ionization-mass spectrometry (LC/ESI-MS). In-tube SPME is an extraction technique for organic compounds in aqueous samples, in which analytes are extracted from a sample directly into an open tubular capillary by repeated draw/eject cycles of the sample solution. Simple mass spectra with strong signals corresponding to [M-H]- and [M-RCOOH]- were observed for all herbicides tested in this study. The best separation of these compounds was obtained with a C18 column using linear gradient elution with a mobile phase of acetonitrile-water containing 5 mmol l-1 dibutylamine acetate (DBA). To optimize the extraction of herbicides, several in-tube SPME parameters were examined. The optimum extraction conditions were 25 draw/eject cycles of 30 microliters of sample in 0.2% formic acid (pH 2) at a flow rate of 200 microliters min-1 using a DB-WAX capillary. The herbicides extracted by the capillary were easily desorbed by 10 microliters acetonitrile. Using in-tube SPME-LC/ESI-MS with time-scheduled selected ion monitoring, the calibration curves of herbicides were linear in the range 0.05-50 ng ml-1 with correlation coefficients above 0.999. This method was successfully applied to the analysis of river water samples without interference peaks. The limit of quantification was in the range 0.02-0.06 ng ml-1 and the limit of detection (S/N = 3) was in the range 0.005-0.03 ng ml-1. The repeatability and reproducibility were in the range 2.5-4.1% and 6.2-9.1%, respectively.

Chromatography, Liquid↗

PubMed2001

Factor transformation to produce statistics describing the uncertainty of analytical data.

Analytical uncertainty produced by random error has a positively skewed distribution and accuracy and precision have non-linear scales. Compared with conventional statistics, factor transformation of the data allows more appropriate interpretation of results and facilitates graphical inspection of data. Statistics are compared for practical examples of performance in proficiency tests and of repeatability and reproducibility in collaborative studies. Factor transformation is shown to be applicable to wide ranges of analyte concentration and measurement precision.

Chromatography, Gas↗

PubMed2002

Flow injection determination of readily assimilable nitrogen compounds during vinification.

A flow injection method for the determination of readily assimilable nitrogen (r.a.n.), i.e. ammonium and aminated nitrogen, is reported. The difference in pH of the sample in the presence and absence of formaldehyde, which blocks the amino function, provides the value of r.a.n. by monitoring the changes in absorbance of bromothymol blue at 616 nm. The detection and quantification limits are 10 and 11.6 mg l(-1), respectively, the reproducibility and repeatability are 3.94 and 1.35 mg l(-1), respectively, and the sample throughput is 20 samples h(-1). The method has been applied to the analysis of 120 samples of must and wine subjected to biological aging. The proposed method also provides good correlation with the reference method used in routine analysis, and is faster

Journal Article↗

PubMed1991

Purification and properties of a neurotensin-degrading endopeptidase from pig brain.

Neurotensin (NT) endopeptidase (EC 3.4.24.16) has been purified about 800-fold from pig brain by four sequential chromatographic steps depending on ion-exchange and hydrophobic interactions. Two types of preparation were studied: one from a Triton X-100-solubilized membrane fraction, and the other from the soluble fraction containing 90% or more of the total activity in the homogenate. NT endopeptidase activity was monitored by high-precision liquid chromatography of the two peptide products, characterized as NT-(1-10) and NT-(1-8), resulting from cleavage of the Pro10-Tyr11 and Arg8-Arg9 bonds respectively. As purification proceeded, from both membranes and cytosol, the yield of the two products achieved a constant ratio of 5:1 and this ratio was reproduced in repeated purifications. However, a distinct peptidase which hydrolysed exclusively at the Arg8-Arg9 bond was partially resolved from NT endopeptidase by chromatography on hydroxyapatite, and this activity was further purified and assigned to endopeptidase-24.15 (EC 3.4.24.15). SDS/PAGE of both preparations of neurotensin endopeptidase revealed a major band of apparent Mr 75000, and treatment of the membrane-associated form with N-Glycanase gave no evidence that the enzyme was a glycoprotein. The membrane-associated and cytosol forms of NT endopeptidase activities, monitored for both NT-(1-10) and NT-(1-8) products, were compared in their responses to 1,10-phenanthroline, EDTA, dithiothreitol (DTT) and some synthetic site-directed inhibitors of endopeptidase-24.15 or peptidyl dipeptidase A. The effects revealed no significant differences between the two preparations, nor did the reagents discriminate between the activities generating the two NT fragments. The partially purified form of endopeptidase-24.15 was also included in this comparison: while some responses were similar, this peptidase was distinguishable in its activation by DTT and its relative resistance to inhibition by EDTA. Both forms of NT endopeptidase were found to hydrolyse other substrates, including Boc-Phe-Ala-Ala-Phe-4-aminobenzoate, bradykinin and substance P (these at faster rates than neurotensin), as well as dynorphin A-(1-8) and luliberin. The bonds hydrolysed in these neuropeptides, as well as in angiotensins I and II and alpha-neoendorphin, were defined. These studies confirm that NT endopeptidase is distinct from endopeptidase-24.15. They further show that the former is a soluble enzyme, not an integral membrane protein, that it is not peptide-specific and that it might be more appropriately named. enzyme, not an integral membrane protein, that it is not peptide-specific and

Amino Acid Sequence↗

PubMed2004

Volkensin from Adenia volkensii Harms (kilyambiti plant), a type 2 ribosome-inactivating protein.

Volkensin, a type 2 ribosome-inactivating protein from the roots of Adenia volkensii Harms (kilyambiti plant) was characterized both at the protein and nucleotide level by direct amino acid sequencing and cloning of the gene encoding the protein. Gene sequence analysis revealed that volkensin is encoded by a 1569-bp ORF (523 amino acid residues) without introns, with an internal linker sequence of 45 bp. Differences in residues present at several sequence positions (reproduced after repeated protein sequence analyses), with respect to the gene sequence, suggest several isoforms for the volkensin A-chain. Based on the crystallographic coordinates of ricin, which shares a high sequence identity with volkensin, a molecular model of volkensin was obtained. The 3D model suggests that the amino acid residues of the active site of the ricin A-chain are conserved at identical spatial positions, including Ser203, a novel amino acid residue found to be conserved in all known ribosome-inactivating proteins. The sugar binding site 1 of the ricin B-chain is also conserved in the volkensin B-chain, whilst in binding site 2, His246 replaces Tyr248. Native volkensin contains two free cysteinyl residues out of 14 derived from the gene sequence, thus suggesting a further disulphide bridge in the B chain, in addition to the inter- and intrachain disulphide bond pattern common to other type 2 ribosome-inactivating proteins.

Amino Acid Sequence↗

PubMed1998

The use of near-infrared spectroscopy for assessing flap viability during reconstructive surgery.

The ability to assess viability of tissues by monitoring changes in oxygenation and perfusion during harvesting and following transfer of free and pedicled flaps is potentially important in reconstructive surgery. Rapid detection of a critical change in tissue oxygenation could enable earlier and more successful surgical intervention when such problems arise. In this study near infra-red spectroscopy (NIRS) was used to assess changes in tissue oxygenation, haemoglobin oxygenation and blood volume in a porcine prefabricated myocutaneous flap model in response to pedicle manipulations. As far as we are aware this is the first usage of a NIRS instrument to assess changes in oxygenation in a flap model which closely simulates the clinical situation. A myocutaneous flap was raised (n = 9 pigs), tubed and the flap circulation allowed to readjust for periods between 7 and 9 days. The pedicle vessels were then subjected to arterial (n = 9), venous (n = 12) and total occlusion (n = 6). Repeatable and reproducible patterns of change were measured in each case. Comparison of mean values indicated that the differences between arterial and venous, and venous and total occlusions were significant for all NIRS parameters. The monitor was easily able to detect two additional features: (i) the presence of venous congestion indicated by raised levels of deoxygenated haemoglobin and an increase in blood volume; and (ii) the presence and magnitude of reactive hyperaemia. In two flaps release of arterial or total occlusion did not result in the expected reactive hyperaemia associated with an increase in blood volume (oxygenated haemoglobin) suggestive of possible damage to the vascular bed. NIRS proved able to detect and distinguish between microcirculatory changes occurring as a result of arterial, venous or total vascular occlusion. We believe that NIRS provides a sensitive and reliable postoperative monitor of tissue viability following transfer of free and pedicled flaps. It can accurately identify different types of problems with the pedicle vessels. In addition its predictive capabilities would allow assessment of flaps buried deep to the skin. This monitor is excellent for surgical and intensive care unit monitoring since it is unaffected by light, portable and is extremely easy to use.

Animals↗