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PubMed1980

Computer densitometry of retinal nerve fibre atrophy. A pilot study.

Lack of nerve fibre layer opacity and a granular appearance of the retina are important funduscopic signs in optic atrophy. In this pilot study serially obtained fundus photographs from a case with developing optic atrophy where subjected to computer densitometry. A monotonic increase in local density variations was found during evolution of optic atrophy. The change in density variations closely followed the funduscopic change in nerve fibre layer opacity. This method makes it possible to detect and quantify diffuse atrophy of the retinal nerve fibre layer.

Adult↗
PubMedDate not supplied

Geometrical considerations for Compton scatter densitometry.

The determination of the physical density by Compton scatter densitometry is dependent upon the geometry used for the measurement. The diameter of the sample, the density range, and the scattering volume size and shape influence the relative electron density values obtained by such a system. This work presents the results of an experimental analysis of these parameters in which the importance of each is evaluated for samples of known density. A bias in the computed Compton density, due to multiple scattering and attenuation, is defined; and an expression relating the density bias to these geometrical parameters is presented. The importance of applying corrections to the computed physical density for samples of large diameter and/or high density (cortical bone tissue and large diameter cancellous bone tissue) is discussed.

Bone and Bones↗
PubMedDate not supplied

Regional silver content of radiographic film determined by x-ray fluorescence compared with optical densitometry.

The regional silver content of radiographic film measured by x-ray fluorescence is compared to light absorption measured by a densitometer. Silver content analysis appears to permit a greater dynamic range of useful exposure levels than does light absorption densitometry. This improvement in latitude, however, is not considered great enough to warrant development of a complex system for scanning silver distribution in radiographic applications.

Densitometry↗
PubMed1993

Estimation of ruminal bacteriophage numbers by pulsed-field gel electrophoresis and laser densitometry.

To investigate phage activity in the rumen, a method for quantifying phage has been developed. By differential centrifugation and ultrafiltration, phage particles were separated and concentrated from ruminal fluid. Linear double-stranded DNA from this fraction containing predominantly tailed phage was isolated and separated by size, using pulsed-field gel electrophoresis (PFGE). Laser densitometry of gel photographs allowed the numbers of phages with DNA in each size region to be calculated and, therefore, the total numbers per milliliter of ruminal fluid to be estimated. Phage numbers were estimated to be between 3 x 10(9) and 1.6 x 10(10) particles ml of ruminal fluid-1. The phage population, as gauged by the appearance of DNA on PFGE gels, had two major components. A broad region of DNA between 30 and 200 kb was always present on PFGE gels. It appears this region comprises DNA from a great many different phages and would include most of the temperate phages. In addition, discrete DNA bands ranging in size from 10 to 850 kb were frequently observed. DNA from one such band, of 12 kb in size, was shown to consist primarily of a single DNA type, suggesting that it originated from a specific phage. It is postulated that the discrete bands are due to epidemics or blooms of phage activity from specific, probably lytic, phages. The method that has been developed will greatly enhance future investigations into the interactions between the ruminal phage population, the ruminal bacterial population, and animal nutrition and growth.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Quantification of specific antibody response to Cryptosporidium antigens by laser densitometry.

Cryptosporidium spp. is a protozoan parasite with worldwide distribution associated with diarrhea in immunocompromised patients (particularly those with acquired immunodeficiency syndrome [AIDS]) and in immunocompetent humans. Immunoglobulin M (IgM) and IgG antibody responses are readily detected by an enzyme-linked immunosorbent assay. To determine which Cryptosporidium antigens invoke antibody responses in humans, we performed polyacrylamide gel electrophoresis using purified oocysts, followed by Western blots with human sera from various populations. Of 40 sera from persons with cryptosporidiosis (24 AIDS and 16 non-AIDS patients), in 37 (93%) a 23,000-dalton antigen measured quantitatively by laser densitometry was recognized. Of 63 sera from IgM- or IgG-positive individuals, as determined by enzyme-linked immunosorbent assay, in 58 (92%) this same antigen was recognized. Up to three additional bands between 125,000 and 175,000 daltons were identified by some of these sera. These results suggest that most persons infected with Cryptosporidium spp. produce antibodies which recognize at least one common low-molecular-weight antigen. Isolation of this antigen will be useful in development of diagnostic tests and may be important in the study of immunity.

Acquired Immunodeficiency Syndrome↗
PubMed1990

Quantitation of antibody reactivity to human immunodeficiency virus (type 1) proteins and glycoproteins on Western immunoblots by reflectance densitometry.

The human serum antibody response to polypeptides of human immunodeficiency virus type 1 (HIV-1) was quantitated by reflectance densitometry of Western immunoblots by using two commercially available blotting systems. In one system, human antibodies were detected by an avidin-biotin method using peroxidase as the label, and in the other, human antibodies were detected by peroxidase-labeled conjugate against human immunoglobulins. When staining intensity was plotted against the log of the serum dilution, a shallow slope was evident, with a 50% change in staining intensity requiring as much as a 100-fold change in antibody content. The linear range of the staining intensity curves was frequently found in serum dilutions of 1:2,500 to 1:1,000,000, and a plateau was often observed at high antibody concentrations (1:80 to 1:640). When replicate strips were tested, staining intensities varied by +/- 7 to 37%. Antibodies to p24gag and gp160env were readily detectable in several sera diluted 1:1,000,000, a result seen with both blotting systems. If Western blotting were to be used to observe increase or decreases in levels of antibodies to various polypeptides, several widely spaced serum dilutions would need to be tested.

Blotting, Western↗
PubMed2003

Performance of clinical referral criteria for bone densitometry in patients under 65 years of age assessed by spine bone mineral density.

BACKGROUND: A case finding strategy based on a number of established risk factors has been suggested by Royal College of Physicians' (RCP) guidelines to optimise bone densitometry referrals for assessment of osteoporosis. OBJECTIVE: The performance of clinical referral criteria was examined in women and men aged <65 years referred for bone mineral density (BMD) assessment. STUDY DESIGN: Cross sectional observational study over six months. RESULTS: Though BMD tended to be lower in patients with multiple criteria for referral, differences from those referred with a single criterion were not statistically significant. The overall prevalence of osteoporosis was higher than expected in both sexes, 11.6% in women and 27.5% in men (expected prevalences were 8% and <1% respectively). BMD was significantly lower in patients referred with a single criterion compatible with the RCP guidelines than in age matched controls or in those patients referred with non-RCP criteria (mean (SD) Z score -0.47(1.38) v 0.35(1.41), p<0.001). Low body mass index was also significantly associated with a lower than expected BMD. In contrast, spine BMD was higher than expected in those with self reported back pain, loss of height, or spinal curvature (p = NS). CONCLUSION: Most of the criteria recommended by the RCP performed well in identifying relatively younger patients with low BMD and osteoporosis. However, prior fractures and corticosteroid use did not reach statistical significance probably due to inclusion of all energy fractures, and current or past steroid use of unspecified dose or duration. Criteria like loss of height and/or spine curvature perform relatively poorly, reflecting the need for further investigation to better identify those needing BMD assessment.

Adult↗
PubMed1996

Quantitative analysis of the IgG and IgG subclass immune responses to chromosomal Pseudomonas aeruginosa beta-lactamase in serum from patients with cystic fibrosis by western blotting and laser scanning densitometry.

BACKGROUND: Antibodies against chromosomal beta-lactamase of Pseudomonas aeruginosa (a beta ab) are markers of the development of resistance of P aeruginosa to beta-lactam antibiotics in patients with cystic fibrosis and chronic lung infection. The role of these antibodies in patients with chronic lung infection with P aeruginosa was further investigated by correlating the a beta ab IgG subclasses with pulmonary function in patients with cystic fibrosis. METHODS: Immunoglobulin G (IgG) and IgG subclass a beta ab were investigated by western blotting and quantified by laser scanning densitometry. A longitudinal study on 43 consecutive patients with cystic fibrosis who developed chronic lung infection with P aeruginosa was performed. RESULTS: IgG subclass a beta ab appeared in all patients with chronic infection with P aeruginosa. Eleven years after the onset of infection all the patients had IgG1, 79% had IgG4, 56% IgG2, and only 16% of the patients had IgG3 a beta ab. The IgG1 and IgG4 a beta ab appeared first, and more than 50% of the patients were IgG1 and IgG4 a beta ab positive within 2-3 years of the onset of infection, but IgG2 positivity only appeared after seven years and IgG3 remained absent from most of the patients. The median a beta ab levels increased during chronic infection: 100-fold for IgG1, 22-fold for IgG2, and 45-fold for IgG4. A 16-fold increase in the IgG3 a beta ab levels was detected in the six patients who developed IgG3 a beta ab. In the first four years of the chronic infection the a beta ab titres were higher in patients with good lung function than in those with poor lung function. CONCLUSIONS: The association of a weak IgG3 and a strong IgG4 a beta ab response suggests that the contribution of a beta ab antibodies to lung diseases mediated by immune complexes might be less important than other antipseudomonal antibodies. A beneficial neutralising effect of the a beta ab antibodies on the antibiotic destroying enzymes may be an additional factor.

Adolescent↗
PubMed1986

Continuous blood densitometry: fluid shifts after graded hemorrhage in animals.

To evaluate rapid fluid shifts after graded hemorrhage in splenectomized animals, four pigs and two dogs were bled 15-23 ml/kg body wt in steps of 2.2-6.0 ml/kg. Arterial blood density (BD), mean arterial pressure (MAP), and central venous pressure (CVP) were recorded continuously, and arterial plasma density (PD) and hematocrit (Hct) were determined from blood samples. Erythrocyte density was computed from PD, BD, and Hct. Starting with stable control conditions, MAP, CVP, and BD fell from the beginning of hemorrhage. Each blood withdrawal was followed by an immediate and rapid decrease in BD, even at the lowest (less than 3 ml/kg) initial blood losses. The time course of BD change mirrored that of the volume replacement, with time constants of 3.0-9.6 min and amplitudes depending on the magnitude of the relative volume loss. The PD decrease was significant (P less than 0.01) after 5.4 +/- 0.7 ml/kg hemorrhage. At 15 ml/kg blood loss the mean PD and BD had dropped by 0.99 +/- 0.15 and 2.42 +/- 0.26 g/l, respectively, and Hct had dropped by 2.40 +/- 0.47 units. Calculations suggest that either the inward-shifted fluid has a higher density than normal ultrafiltrate and/or there is a rise of the whole-body-to-large vessel Hct (F cell ratio). The rapid fluid replacement ranged from 5.8 +/- 0.8 to 10.6 +/- 2.0% of the initial plasma volume, or one-fifth to one-third of the lost volume with a 20% hemorrhage. Transvascular fluid shifts can be monitored with continuous high-precision blood densitometry.

Animals↗
PubMed1986

Method of high-precision microsample blood and plasma mass densitometry.

The reliability of the mechanical oscillator technique (MOT) for blood and plasma mass density measurements on small samples is quantified in this paper. Sources of measurement errors that can reduce both the accuracy and precision of density determinations include storage of plasma samples, inhomogeneity of blood samples, and density reading before adequate temperature equilibration. Measurements on fractions from identical samples and repeated samplings from test subjects under steady-state conditions revealed a 10(-2) g/l reproducibility of density readings. The mean plasma density (PD) readings did not change significantly after up to 1-wk storage at +4 degrees C or up to 2 mo storage at -20 degrees C. The variability of the PD findings increased with storage time and were generally higher with storage at -20 degrees C, compared with +4 degrees C. Densitometers of different sizes were used to evaluate rheological influences on blood density (BD) readings. Linear correlations between PD and plasma protein concentration, between BD and blood hemoglobin concentration, and between erythrocyte density and mean corpuscular hemoglobin concentration were significant (P less than 0.001). Rapid density measurements with up to 10(-2) g/l reliability on small (less than 0.1 ml) volumes of biological fluids and continuous blood densitometry can be performed with use of the MOT.

Animals↗
PubMed1993

Capillary filtration coefficients using laser densitometry and gravimetry in isolated dog lungs.

We compared pulmonary capillary filtration coefficients (Kf,c) using measurements of transcapillary filtration rates based on laser densitometry of perfusate hematocrit changes (Jy,l) and gravimetric measurement of the rate of lobe weight gain (delta Wt/delta t) after an increase in capillary pressure (Pc) in isolated autologous blood-perfused dog lungs. Although the lobe weight increased at a rate that decreased with time, the densitometric filtration rate was relatively constant over time. Kf,c values were calculated in milliliters per minute per centimeter water per 100 g from filtration rates obtained by 1) extrapolation of the delta Wt/delta t from 3 to 10 min back to time 0 [Kf,c(0)], 2) use of the slope of the line fitted to the delta Wt/delta t and Jv,l values obtained after three stepwise increases in Pc [Kf,c(slope)], and 3) use of delta Wt/delta t and Jv,l values obtained 10, 20, and 30 min after either a constant 30-min or three 10-min stepwise increases in Pc [Kf,c(t)]. The mean Kf,c(0) values ranged from 0.171 +/- 0.024 to 0.188 +/- 0.070 and were not significantly different between vascular pressure states. These Kf,c(0) values were significantly higher than all gravimetric or densitometric Kf,c(t) or Kf,c(slope) values that ranged from 0.037 to 0.100. Kf,c(t) values were not statistically different from each other by use of either method, but the gravimetric Kf,c(slope) was significantly higher than the densitometric value. Therefore, extrapolated Kf,c(0) probably overestimates endothelial barrier Kf,c because of persistent vascular stress relaxation after a vascular pressure increase. Adjustment of tissue Starling forces may occur rapidly after an increase in filtration pressure.

Animals↗
PubMed2006

Determination of berberine content in the stem extracts of Coscinium fenestratum by TLC densitometry.

OBJECTIVE: To develop the optimal extraction procedure (i.e. maceration, percolation or Soxhlet extraction) and thin- layer chromatographic (TLC)-densitometric method for the determination of berberine content of Coscinium fenestratum. MATERIALS AND METHODS: Maceration, percolation and Soxhlet extraction techniques were used to extract alkaloids from dried stems of C. fenestratum. The solvents used were 50 and 80% ethanol. Crude extracts and berberine content recovered from the TLC fingerprint were evaluated for chemical components of each extraction method. Precoated silica gel GF(254) plates were used as stationary phase while butanol:glacial acetic acid:water (14:3:4) was used as a mobile phase. Detection and quantitation of berberine were performed by densitometry at the wavelength of 415 nm over the linearity range of 240-840 ng (r(2) = 0.9982). The relative standard deviations from intraday and interday precisions were less than 4.13%. RESULTS: The recovery of standard berberine was 97.58-98.71% (%RSD = 3.85), and the limit of detection and quantitation were 25 and 50 ng/spot, respectively. Eighty percent ethanol gave a higher content of berberine than 50% ethanol. Berberine contents from maceration, percolation and Soxhlet extraction with 80% ethanol were 3.37+/- 0.30, 3.08+/- 0.38 and 2.67+/- 0.27% w/w, respectively. CONCLUSION: The TLC-densitometric method was simple, accurate and precise for quantitating berberine in the stem extract of C. fenestratum. Maceration with 80% ethanol gave the highest content of berberine in the extract. TLC of the extracts from different methods showed a similar pattern.

Berberine↗
PubMed2007

Quantitative analysis of barakol content in Senna siamea leaves and flowers by TLC-densitometry.

OBJECTIVE: To develop a TLC-densitometric method for the determination of barakol content in Senna siamea leaf and flower extracts, and to compare the barakol content in mature leaves, young leaves and young flowers of the plant which are consumed as a vegetable in curry. MATERIALS AND METHODS: The extraction of pure barakol was performed by boiling the fresh young leaves of S. siamea with 0.5% sulfuric acid followed by chloroform extraction. The extract was further purified and recrystallized from absolute ethanol. Authentic sample of barakol was used for the validation of the TLC-densitometric method. Chromatography was performed on a TLC aluminium plate precoated with silica gel 60 F(254)as a stationary phase and chloroform-methanol (85:15 v/v) as a solvent system. Fifteen percent ethanolic extracts of mature leaves, young leaves and flowers of S. siamea were analyzed and compared for barakol content using the validated TLC-densitometric method. Both the validation and analysis of barakol by TLC-densitometry were carried out at the absorbance mode of 366 nm. RESULTS: Barakol was extracted as pure lemon-yellow crystals from young S. siamea leaves with 0.1% yield. Linearity was found over the range of 200-900 ng/spot (r(2) = 0.997). The developed method gave high precision (%RSD < 0.50) and accuracy (average 101.12%). The limit of detection and limit of quantitation were 8 and 50 ng, respectively. Barakol content in young leaves, mature leaves and young flowers were 1.67, 0.78 and 1.43% dry weight, respectively. R(f) value of the barakol in young leaves, young flowers and authentic sample was the same: 0.45 +/- 0.03. CONCLUSION: The TLC-densitometric method was simple, precise and convenient; hence it is an effective procedure for the simultaneous determination of barakol in plant extracts.

Benzopyrans↗
PubMed1996

High-resolution computed tomographic evaluation of the cochlear capsule in otosclerosis: relationship between densitometry and sensorineural hearing loss.

Otosclerotic cochlear involvement is a rather frequent disease that has not been clearly understood in terms of diagnosis and management. Objective evaluation methods are needed to confirm the clinical diagnosis, investigate the relationship with hearing impairment, and validate the results of treatment. In this study two ear groups with bone conduction hearing loss (BCHL) were investigated with audiometry and high-resolution computed tomography (HRCT). In the first group (n = 22) the diagnosis of fenestral otosclerosis was confirmed at operation; the second group (n = 9) was composed of ears clinically suspicious for purely cochlear involvement. Additionally, a control group (n = 14) of otologically normal ears was also studied. Foci of demineralization were demonstrated in 58% of the ears in the two groups; the sensorineural hearing loss (SNHL) in those ears was significantly worse than in those with normal radiologic findings. Three methods of HRCT densitometry were used to determine the abnormal regions in the cochlear capsule; the results suggested that hypodense regions were consistent with a greater degree of SNHL, in contrast to the hyperdense ones in ears with better cochlear reserves. Agreement was found between the location of the density change and the frequency topography of the SNHL; densitometric values were correlated with the bone conduction thresholds for certain frequencies. It is concluded that the spongiotic foci are responsible for the SNHL, since there was a correlation between their location and the SNHL frequency. The determination of better hearing in those ears with sclerotic foci supports the hypothesis that the sclerotic phase may not be a healing process following the spongiotic phase, and that it can be the first stage of the disease.

Cochlea↗
PubMed1984

Analysis of individual bile acids and their glycine/taurine conjugates by high-performance thin-layer chromatography and densitometry.

We have investigated a thin-layer chromatography method for the quantitative measurement of free and conjugated bile acids in duodenal juice. Evaluation of solvent systems and colour spray reagents has enabled us to select conditions which improve the resolution, sensitivity and colour stability of the method. The modified method, using high-performance thin-layer chromatography and densitometry, is rapid, accurate, reproducible and practicable. The technique deserves wider routine use.

Bile Acids and Salts↗
PubMed1990

Estimation of paraproteins by immunoturbidimetry and electrophoresis followed by scanning densitometry.

A comparison of paraprotein estimation by immunoturbidimetry (IT) and electrophoresis followed by scanning densitometry (EFD) showed significant differences in concentration for IgG, IgA, and IgM paraproteins. Overestimation of IgM paraproteins by IT was particularly marked. Our data suggest that both techniques should be used for IgG and IgA paraproteins but for IgM paraproteins IT was found to be unsuitable and EFD is the method of choice.

Blood Protein Electrophoresis↗
PubMed1987

Localization and characterization of insulin receptors in rat brain and pituitary gland using in vitro autoradiography and computerized densitometry.

In order to identify likely sites of action in insulin in rat brain we have used the technique of in vitro autoradiography and computerized densitometry to map, characterize, and quantify its receptors in coronal and sagittal sections. A discrete and characteristic distribution of insulin receptor binding was demonstrated, with specific binding representing 92% of total binding. Displacement and specificity competition curves in olfactory bulb are typical for authentic insulin receptors, and computer analysis indicates a single class of binding site with a dissociation constant (Kd) 0.48 nM for choroid plexus and 0.44 nM for olfactory bulb external plexiform layer. Insulin receptor density is maximum in the choroid plexus, and high in the external plexiform layer of olfactory bulb. Structures of the limbic system and hypothalamus reveal moderate to high insulin receptor density, particularly the lateral septum, amygdala, subiculum, hippocampal CA1 region, mammillary body, and arcuate nucleus. Moderate insulin receptor density occurs in regions of cerebral cortex and cerebellum, and moderate to low binding occurs in discrete brainstem and midbrain structures. Insulin binding in the pituitary gland is greatest in the anterior lobe, with clear distinction from intermediate and posterior lobes. The circumventricular organs and the thalamus show low insulin binding. We conclude that insulin receptors are widespread throughout rat brain, with concentration in regions concerned with olfaction, appetite, and autonomic functions. The distribution is distinct from other neuropeptides and not related to either vascularity or cell density. A common feature of regions rich in insulin receptors is that they contain dendritic fields receiving rich synaptic input. Whether insulin plays a specific neurotransmitter or metabolic role in these sites remains unclear, but these studies have provided detailed information on potential sites of action of insulin in the brain, and will allow further studies to examine insulin receptor function in specific brain regions.

Animals↗
PubMed1990

21-hydroxylase deficiency: disease-causing mutations categorized by densitometry of 21-hydroxylase-specific deoxyribonucleic acid fragments.

The types of disease-causing mutations were studied in 43 unrelated patients with 21-hydroxylase deficiency. Densitometry of Southern blots after cleavage with the restriction enzymes TaqI, PvuII, and BglII was used to measure the ratio of the copy-number of the 21-hydroxylase gene (CYP21) to the copy-number of its pseudogene (CYP21P). DNA from 16 unrelated patients showed equal hybridization intensities of the 2 genes, indicating that point mutations caused the enzyme deficiency. One of the 2 haplotypes in 7 patients showed evidence of a large gene conversion between the CYP21 and the CYP21P gene without loss of the total number of 21-hydroxylase genes. Deletion of at least 1 21-hydroxylase gene was found in 11 patients. DNA from 8 of these patients had relative hybridization intensities compatible with a deletion of the active 21-hydroxylase gene, CYP21. Two patients with the salt-wasting form of the disease showed homozygous loss of DNA fragments that are specific for the 5' end of the active 21-hydroxylase gene. Nine patients showed relative 21-hydroxylase hybridization intensities compatible with duplication of the gene in 1 or both haplotypes. In conclusion, point mutations, gene conversions, or CYP21 gene deletions are the typical mutations in patients with the simple virilizing and salt-wasting forms of the disease, while duplications of the locus are overrepresented in patients with nonclassical 21-hydroxylase deficiency.

Adrenal Hyperplasia, Congenital↗