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PubMed1981

The leukocyte left shift in clinical and experimental neonatal sepsis.

The leukocyte left shift is commonly used as an adjunct to the early diagnosis of bacterial infection. Many different methods have been employed for its quantification, such as the absolute band count, band/seg ratio, band/total neutrophil ratio, and immature/total neutrophil ratio. We examined blood and bone marrow samples in groups of noninfected and infected neonatal dogs and human beings in order to determine which method most clearly reflects an increased call upon marrow neutrophil reserves and which correlates best with the presence and severity of infection. We found that the neutrophil ratios were more frequently abnormal during neonatal sepsis than was the the absolute band count. All subjects, canine and human, in whom the immature/total neutrophil ratio exceeded 0.800 were found to have depletion of the marrow neutrophil reserves, and those with the most profound depletion died. This study supports the concept that an elevated immature/total neutrophil ratio can aid in the diagnosis of bacterial infection in the newborn infant and suggests that the degree of elevation may serve as a method for detecting subjects at high risk for depletion of the marrow neutrophil reserves and death from sepsis.

Animals↗

PubMed2026

Telomere length in patients with non-functional adrenal incidentalomas.

Telomeres maintain genomic integrity during cell replication by preventing chromosomal fusions. Beside genetic influences, telomere length is affected by environmental factors such as oxidative stress and inflammation. These mechanisms also contribute to metabolic syndrome components linked to cellular aging. We aim to evaluate whether telomere length is shortened in patients with non-functional adrenal incidentaloma (NFAI) compared to the control group. This study was designed as a prospective, single-center study. The total of 88 participants included were 44 patients aged between 40 and 60 years with NFAI in our endocrinology clinic and 44 control subjects. An Absolute Human Telomere Lengths Quantification qPCR Assay kit (Nucleotestbio, Budapest, Hungary) was used for analyses. There was no significant difference between the NFAI and control groups regarding age and sex distribution. Telomere length was significantly shorter in the NFAI group (NFAI group: 3.680 ± 1.970 kb; control group: 4.469 ± 1.672 kb; p = 0.046). While no significant difference was found in telomere lengths in subgroup analyses, patients with basal adrenocorticotropic hormone (ACTH) levels <15 pg/mL had significantly shorter telomeres than those with basal ACTH levels ≥15 pg/mL (p = 0.034). A strong positive correlation was observed only between telomere length and ACTH level (p = 0.001). This study demonstrated that telomere length is significantly shortened in NFAI patients. Here, we propose that the underlying cause of telomere length shortening in the NFAI group may be related to increased cardiovascular risk and an elevated inflammatory state, even in the presence of cortisol levels within the normal range.

Humans↗

PubMed1993

Absolute concentrations of metabolites in the adult human brain in vivo: quantification of localized proton MR spectra.

In vivo concentrations of cerebral metabolites were obtained by means of 52 single-voxel, localized proton magnetic resonance (MR) spectroscopic examinations of different regions of the brain performed in 26 healthy adults aged 21-32 years. The study was performed at 2.0 T with use of a circularly polarized head coil to ensure homogeneous radio-frequency excitation and signal reception. Proton MR spectra were obtained in the stimulated-echo acquisition mode under fully relaxed conditions (repetition time > or = 6,000 msec) and at short echo times (20 msec) to minimize corrections due to T1 and T2 attenuation and depict the spectra of metabolites with strongly coupled resonances. Absolute concentrations were obtained by means of calibration of resonance signal areas with those of pertinent metabolite solutions from separate studies and correction for coil loading and partial volume effects (eg, with perfused capillary networks and cerebrospinal fluid). The results provide a quantitative basis for studies of both normal human neurochemistry in vivo and metabolic alterations in diseases of the brain.

Adult↗

PubMed1994

A new method of quantification of the pulmonary regional distribution of aerosols using combined CT and SPECT and its application to nedocromil sodium administered by metered dose inhaler.

Aerosols of nedocromil sodium labelled with 99Tcm were delivered on 20 separate occasions to healthy male volunteers. Planar and single photon emission computerized tomography (SPECT) gamma scintigraphy were immediately performed to assess the pulmonary regional distribution of delivered aerosol. On a separate occasion volunteers were imaged using X-ray computed tomography (CT). Alignment of SPECT and CT images was performed using marked anatomical features and the anterior and lateral skin outlines. CT images provided data for attenuation correction and were used to define the anatomical lung volume. Central to peripheral (CP) ratios of deposited activity were calculated from volumes of interest in coronal and transverse sections of the right lung. These were compared with CP ratios obtained from planar images obtained immediately following aerosol inhalation. Volumetric CP ratio correlated significantly with immediate planar CP ratio (p < 0.001). Analysis of deposition in the whole right lung was performed by separating the SPECT lung data into a series of thin concentric shells centred on the entry of the right main bronchus. Measures were defined for describing the variation of deposition density and cumulative total deposition with distance from the lung centre. These showed significant correlation with planar CP ratio (p < 0.001). SPECT analysis using CT is consistent with planar measures of aerosol deposition but offers a more complete quantification of aerosol penetration and absolute deposited activity within the whole lung. It is a valuable new tool for aerosol analysis.

Administration, Inhalation↗

PubMed1995

[Value of various suture materials for defect reconstruction of peripheral nerves].

A total of 40 surgical procedures were performed for bridging defects of the peronaeus nerve in 20 experimental animals (White Russian rabbits) with absorbable and non-absorbable suture materials. To investigate whether differences in the biological behaviour of suture materials may influence the formation of axons in the coaptation areas positively or negatively, the nerves were examined histologically and electrophysiologically and the regeneration process in the m. tibialis anterior, the reacting organ of the n. peronaeus, was assessed by histomorphometric quantification. Despite minor deviations in absolute values, significance tests and statistical evaluation with Wilcoxon's test revealed no significant differences among the groups. We could show that differences in biological behaviour of absorbable and non-absorbable suture materials are without influence on nerve regeneration in the proximal and distal coaptation areas and therefore without effect on the final results of nerve regeneration in defect areas.

Animals↗

PubMed1995

Imaging of the heart by MRI and PET.

This review article describes the clinical usefulness and future potential of two new methods for the imaging of the heart, which have recently also become available outside research laboratories. These methods are magnetic resonance imaging (MRI) and positron emission tomography (PET). MRI is the most rapidly increasing imaging modality in medicine today. The moving heart forms a challenge to conventionally rather slow MRI-techniques. Techniques based on ECG-gating have been mandatory in making cardiac cine-MRI possible. Even though MRI provides accurate and quantitative information of the heart, conventional methods are time-consuming, confined to special laboratories, and rather expensive. Therefore, the clinical use of cardiac MRI is, in many laboratories, limited to cases in which echocardiography does not provide adequate information (e.g. pulmonary circulation) or when the patient is not willing to have transoesophageal echocardiography for better visibility. MRI is also used instead of or to complement invasive angiography to study large vessels, and it provides excellent information on paracardiac masses. Cardiac MRI is developing rapidly and within the next few years it is likely to have a profound impact on cardiac imaging. This is based on its noninvasive nature and on the comprehensive anatomic (including coronary arteries), functional, flow, perfusion and possibly also metabolic information it has the potential to provide in a manner not comparable to any other imaging method. PET is a nuclear medicine imaging modality that allows quantitative characterization of a variety of physiological and metabolic processes in vivo. Using positron-emitting flow tracers and analogues of metabolic substrates, regional myocardial blood flow, glucose and fatty acid metabolism and oxygen consumption can be studied noninvasively by PET in research as well as in clinical practice. For example, regional myocardial glucose utilization rates can be measured accurately by PET. This allows us to study the effects of nutritional interventions, hormonal and neural effects as well as disease processes on the glucose utilization of the human heart. PET is currently the only technique that permits noninvasive quantification of regional myocardial perfusion in absolute terms. Over the last decade, PET has also emerged as a clinically useful tool to study coronary artery disease and myocardial viability.

Heart↗

PubMedDate not supplied

Absolute lung density in experimental canine pulmonary edema.

The noninvasive quantification of pulmonary edema could be of importance in patient management. We have developed a portable Compton-scatter densitometer capable of measuring density in the range of 0.1-1 g/cm3 independently of the chest wall. Change in lung density was investigated in 19 dogs with pulmonary edema induced by a combination of raised left atrial pressure and hemodilution. The increase in lung density correlated with the postmortem assessment of lung water (r = 0.862, p less than 0.01) and with hemodynamic indicators of pulmonary edema (r = 0.749, p less than 0.01). Subtracting the intravascular component of lung density did not improve the correlation with postmortem lung water (r = 0.850, p less than 0.01). The measurement of absolute lung density by a Compton-scatter technique may be a potentially useful method of quantifying pulmonary edema.

Absorptiometry, Photon↗

PubMed2026

Accurate quantification of canine mitochondrial DNA copy number from canine blood and brain samples.

Acute brain injury is difficult to evaluate in veterinary medicine and tools to investigate the potential involvement of mitochondrial involvement are limited. The brain is highly enriched in mitochondria and contains thousands of copies of mitochondrial DNA (mtDNA) per cell, but robust methods for quantifying mitochondrial DNA copy number (mtDNA-CN) in canine tissues are lacking. We describe the development of a quantitative real-time PCR assay for absolute measurement of mtDNA-CN which was validated in canine blood and brain tissue. To minimize amplification of nuclear mitochondrial insertion sequences (NumtS) and repetitive regions, species-specific oligonucleotide primers were designed following in silico genomic filtering. The assay was applied to a small pilot cohort comprising blood samples from dogs with and without acute brain injury (n = 4-6 per group) and cerebral cortex samples (n = 1 per group) to assess feasibility and biological plausibility. In non-brain injury dogs, blood mtDNA-CN ranged from 98 to 288 copies per nuclear genome (mean 193 ± 72), while values in brain-injured cases ranged from 163 to 228 copies per genome (mean 200 ± 33). Cerebral cortex samples exhibited higher mtDNA-CN than blood, consistent with known tissue-specific mitochondrial enrichment. In a single brain-injured case with serial sampling, mtDNA-CN increased over five days. This study presents a validated assay and pilot data for mtDNA-CN quantification in canine samples. While not powered for biomarker evaluation, this method may enable future studies of mitochondrial dynamics in canine brain injury and metabolic disease.

Animals↗

PubMed1994

Early detection of abnormal coronary flow reserve in asymptomatic men at high risk for coronary artery disease using positron emission tomography.

BACKGROUND: The objective of this study was to compare coronary flow reserve (CFR) as a measure of vascular integrity in asymptomatic middle-aged men with family history of coronary artery disease (CAD) and a high-risk lipid profile with men without risk factors for CAD using positron emission tomography (PET). Previous studies suggested that the assessment of CFR is a sensitive means to detect vascular abnormalities before angiographic appearance of CAD. N-13 ammonia PET scanning allows noninvasive evaluation of regional and global myocardial blood flow and thereby quantification of CFR. METHODS AND RESULTS: We used dynamic N-13 ammonia PET imaging in conjunction with intravenous adenosine to assess regional and global CFR in asymptomatic middle-aged men with high risk (group 1, n = 16) and men without any known risk factors (group 2, n = 11) for CAD. Group 1 patients were selected based on positive family history of CAD, one or more lipid abnormalities, and a normal stress test. No patient had history of diabetes or hypertension. A three-compartment tracer kinetic model developed and validated in our institution was used to calculate myocardial blood flow. Absolute myocardial blood flow (mL/100 g per minute) was calculated in five territories for each patient. CFR was defined as the ratio of blood flow during maximum pharmacological vasodilatation to blood flow at rest. Comparisons of CFR between the two groups of patients were performed. The mean age was similar between groups (group 1, 49.3 +/- 0.5 years; group 2, 48.1 +/- 8.7 years; P = NS). Group 1 had higher total cholesterol (mg/dL) (241 +/- 43 versus 173 +/- 34, P < .001), total cholesterol to high-density lipoprotein cholesterol ratio (6.4 +/- 1.6 versus 4.1 +/- 1.4, P < .001), and low-density lipoprotein cholesterol (mg/dL) (167 +/- 33 versus 107 +/- 32). No group 1 patient had evidence of ischemia by exercise ECG or exercise of pharmacological radionuclide perfusion studies. The mean global absolute myocardial blood flow at rest was not significantly different among groups (group 1, 76 +/- 18; group 2, 66 +/- 8; P = NS; (in mL/100 g per minute). However, blood flow after adenosine infusion was higher for group 2 (group 1, 217 +/- 56; group 2, 264 +/- 39; P < .001), which resulted in a larger CFR for group 2 (group 1, 2.93 +/- 0.86; group 2, 4.27 +/- 0.52; P < .001). Univariate linear regression analysis revealed significant negative correlation of CFR to total cholesterol (P < .05, r = -.41), low-density lipoprotein (P < .05, r = -.38), and total cholesterol to high-density lipoprotein cholesterol ratio (P < .05, r = -.47). CONCLUSIONS: Noninvasive quantification of absolute myocardial blood flow by N-13 ammonia PET allows the detection of abnormal vasodilatory response to intravenous adenosine in male patients with family history of CAD and high-risk lipid profiles. Early assessment of alterations of vascular reactivity to adenosine in relation to high-risk lipid profiles in asymptomatic men may allow early detection of preclinical atherosclerosis and may initiate modification and/or elimination of risk factors that may slow, retard, or even reverse the progression of CAD.

Adenosine↗

PubMed1994

Assessment of epidermal dendritic cell markers and T-lymphocytes in psoriasis.

Epidermal dendritic and T-cell counts have been performed in lesional and non-lesional skin from 35 psoriatic patients. The aims were to investigate absolute changes and interrelationships between these cellular elements in psoriasis and to explain apparent discrepancies between these results and reports in the literature. In non-lesional skin, the most frequently expressed dendritic cell marker was CD1a. HLA-DR+ and alpha-mannosidase+ dendritic cells were approximately 50 per cent and S100+ cells were 25 per cent as frequent. T-lymphocytes were rare, CD4+ cells predominating. In lesional psoriatic epidermis, there was a definite increase (approximately two-fold) in the absolute number of CD1a+ dendritic cells. This differs from the conclusions from the majority of previous studies. However, when cell counts were expressed per unit area of vertical section, there was a decrease in CD1a+ cells in lesional skin, which is an explanation for this discrepancy. There was a greater increase in absolute HLA-DR+ cell counts, so that the numbers of cells expressing CD1a and HLA-DR were similar in lesional skin. S100 expression increased proportionately with CD1a+, but there was no absolute increase in alpha-mannosidase+ cells, which might represent a separate sub-population of dendritic cells. The greatest cellular increase was in T-lymphocytes, particularly CD8+. In lesional skin, direct correlations have been demonstrated between epidermal thickness, HLA-DR+ dendritic cells and T-lymphocytes, particularly CD8+ cells. We would suggest that the present method of quantification is of value for the analysis of absolute changes in epidermal infiltrates, particularly psoriasis, and could be applied to other epidermal pathologies.

Adult↗

PubMedDate not supplied

A preliminary phantom study on a proposed model for quantification of renal planar scintigraphy.

A method for estimation of the absolute renal activity within background activity in renal planar scintigraphy will be presented. This method corrects for oversubtraction of background activity with consideration of background activity of the kidney volume, and employs the depth-independent buildup factor (DIBF) method for the correction of attenuation and scatter of photons. This method requires the depth and thickness of the organ and the thickness of background for a background activity correction. The transmission factor (TF) for a volume source is derived from integrating TF for a thin source over the thickness of the organ for the DIBF method. To validate this method, phantom studies with various uniform background activity concentrations were performed and the data were compared with conventional background subtraction that do not consider the organ's volume. The results showed that with the conventional background subtraction method at all depth activity was underestimated with errors of 5%-30% for organ/background concentration ratios of 5-40, while this method estimated the true count rate with errors of less than 5%. More accurate quantifications of renal functions such as renal uptake, glomerular filtration rate (GFR) and renogram may be obtained by this proposed method on the planar images.

Biophysical Phenomena↗

PubMed1995

A synthetic standard DNA construct for use in quantification of murine cytokine mRNA molecules.

A synthetic DNA construct has been developed as a standard molecule whereby murine cytokine mRNA molecules can be quantified by the reverse transcription-polymerase chain reaction (RT-PCR). The construct, designated Cytoquant 1, allows the quantification of murine IL-1 alpha, IL-2, IL-3, IL-4, IL-5, IL-6, IL-10, IFN-gamma, TNF-alpha, TGF-beta, GM-CSF, CD4, CD8, HPRT and beta-actin mRNA levels. This technique is based on the amplification of a transcribed RNA molecule from Cytoquant 1 as an internal standard control in both the RT and PCR reactions. The quantification data from these analyses are expressed in absolute values, i.e. molecules/cell, which allows the data derived from separate experiments to be compared. In this study, mRNAs encoding beta-actin, IL-10, IFN-gamma and GM-CSF have been quantitated in both Th1 and Th2 cell clones with, and without, stimulation. The quantitative analysis data are highly reproducible and cytokine mRNA concentrations are reflective of restricted cytokine secretion patterns. Furthermore, constitutive cytokine mRNA levels are detectable in resting cells, eliminating the need for exogenous stimulation. The high degree of sensitivity and accuracy make this methodology uniquely suited for the study of T-cell subset cytokine expression in both in vivo and in vitro biological models.

Animals↗

PubMed1975

Liquid chromatography in pharmaceutical analysis IV: determination of antispasmodic mixtures.

Parameters associated with the separation of antianxiety-antispasmodic agents were investigated using high-pressure liquid chromatography. Eight widely prescribed drugs were studied. The compounds were chromatographed on reversed-phase octadecyltrichlorosilane (C18) or diphenyldichlorosilane (phenyl) columns, using mixtures of absolute methanol and distilled water buffered with ammonium dihydrogen phosphate, ammonium acid phosphate, or ammonium carbonate. A mixture of phenobarbital-propantheline bromide was selected to demonstrate the utility of the separation and quantification method. The mixture was chromatographed on a phenyl column, using absolute methanol-aqueous 1 percent ammonium dihydrogen phosphate (60:40) (pH 5.85) at a flow rate of 1.4 ml/min. Each determination can be achieved in approximately 15 min with an accuracy of 1-2 percent.

Atropine↗

PubMed1991

[Quantification of T-lymphocyte subpopulations. Immunological marker in human immunodeficiency virus infections].

The absolute number of CD4+ and CD8+ T cells was counted directly in the blood of 75 healthy controls and 223 individuals infected with human immunodeficiency virus (HIV). The HIV-seropositive individuals were also classified clinically according to the system recommended by the Centers for Disease Control. We observed a pattern of changes in the T cell subset counts of the patient group. This pattern could be defined by five T cell stages ranging from normal T cell subset values to values representing severe T cell immunodeficiency. A close correlation was observed between the immunological and clinical classification of the patients. Quantification of T cell subsets may provide useful information in the follow-up of individuals infected with HIV, particularly in assessing the indications for antiviral therapy, and its effects.

HIV Infections↗

PubMed1991

Quantification of glutamate, glutamine, and other metabolites in in vivo proton NMR spectroscopy.

A reliable quantification of in vivo 1H MRS spectra is hampered by extensive line overlap, by intensity distortions due to the water suppressing pulse sequence and complex modulation patterns of J-coupled spins, and by deviations between the experimental and model lineshapes in computer fitting programs. By correcting the experimental lineshape, using a suitable pulse sequence, and incorporating prior knowledge about the spin systems and the intensity distortions, a sufficient quantification is possible. If reliable T2 values are available absolute concentrations can be determined.

Animals↗

PubMed1987

[The effect of terguride on the dynamics and mean level of vigilance. A controlled electroencephalographic study with elderly subjects].

The influence of a 3-week medication period with terguride (trans-dihydro-lisuride) in a dosage of 2 X 0.2 mg/d per os upon the 10-min resting EEG of elderly, clinically normal volunteers (mean age 70 years was investigated. Eight subjects received placebo and 7 subjects received terguride under double-blind conditions. The EEG data were analysed, inter alia, by a newly-developed method of quantification which uses information on topographical relationships of the absolute alpha-powers provided by two anterior and two posterior leads. Under terguride, but not under placebo, a significant increase of subvigilant patterns was found. If one were to disregard the baseline situation, which was characterized in the present sample by a distinct restriction of the physiological dynamics of vigilance at a reduced level (dynamic rigidity), this finding would have to be interpreted as sedation. A non-physiological pre-drug situation was indicated by marked and sustained anteriorization of alpha-activity. Taking into account our additional finding that terguride, but not placebo, eliminates this non-physiological alpha-anteriorization, we interpret the increase of subvigilant patterns in the course of the recording period as a resolution of the preexisting dynamic rigidity and, consequently, as a therapeutically desirable effect.

Aged↗

PubMed1992

High pressure liquid chromatographic analysis of the serum concentration of cefuroxime after an intravenous bolus injection of cefuroxime in patients with a coronary artery bypass grafting.

A simple reversed-phase high pressure liquid chromatographic method was developed for the determination of cefuroxime in the serum of patients undergoing coronary artery bypass grafting. The serum was cleaned up with a 3.3% solution of perchloric acid in water. Cefalexine was used as an internal standard. Detection was made by a UV multi-wavelength detector. The optimum wavelength for cefuroxime is 275 nm. The absolute recovery of this method was 90.9%; the limit of quantification was 0.7 mg/l. This analytical method was used in a study to investigate the cefuroxime serum concentration--time curves in 26 patients undergoing coronary artery bypass grafting. It was found that one single dose is sufficient to obtain effective serum concentrations.

Adult↗

PubMed1986

Quantification of cell surface roughness; a method for studying cell mechanical and adhesive properties.

The presence on the surface of nucleated cells of a variety of asperities of different size and shape plays a prominent role in cell-cell and cell-substrate interaction. Also, the organization of these asperities is directly related to cellular cytoskeletal elements. In the present report, we describe a simple and objective method of studying electron micrographs to quantify the roughness of cell contours. Constant-length segments of cell boundaries are compared to reference circular segments with common extremities and enclosing the same area. This procedure was performed with a digitizer connected to a microcomputer, and it was used to analyse model contours or electron micrographs of (i) target tumour cells bound by cytotoxic T lymphocytes and (ii) thymocytes sticking to concanavalin A-coated surfaces. It is shown that this method allows precise quantification of cell deformation in adhesive zones, which may allow absolute evaluation of adhesive stimuli.

Animals↗