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Biomedical subjects

H Barden

Publications and source records attributed to H Barden.

At least 19 recordsLinked to original sources

Bone density of the spine and femur in adult white females.

We measured bone mineral density (BMD in g/cm(2)) of the spine (L2-L4) and femur (four regions) in 1472 and 1487 cases, respectively, of ambulatory white women ages 20-79 years in the USA. A DPX densitometer was used in a mobile setting. The BMD values for women up to 69 years corresponded closely with published values for the USA, the UK, and northern Europe; our values were somewhat lower than those from other studies only in women over 70 years. The USA data were combined with data from Europe to give reference curves on about 12,000 subjects. Decreases of BMD with age in women below 50 years were much smaller than in older women (0.2% versus 0.6-1.0% per year). Femoral bone decreased from the neck region, but not the trochanter with age; the decrease of total femur BMD with age was due to loss from the former region. Loss of bone mineral content (BMC in g) from the femur neck and total femur region did not accelerate until after age 50 years, much like the spine. The apparent decrease of BMD in these regions that begins about age 40 actually is due to an increase of bone area. About 20% of USA women aged 50-79 years had BMD levels for the lumbar spine, or for the femur neck, more than -2.5 SD below the average values in young adult women 20-39 years old. Body weight had several times more impact on BMD than height, and in fact, a change of 1 kg in postmenopausal women was commensurate with the effect of a 1-year change in age. Subjects in the lowest quartile of body weight had T-scores that were 1 SD below those in the highest quartile.

Absorptiometry, Photon↗

Normalization of spine densitometry.

We investigated several transformations of bone mineral content (BMC) and area density (BMD), in particular volumetric density (BMAD), to ascertain the influence on (1) body size dependence, (2) diagnostic sensitivity, and (3) precision. These transformations were examined in a group of 657 normal postmenopausal women and 327 women with osteoporotic fracture. First, expression of results as BMAD removed some of the slight dependence on body size; 21% of the variation in BMC and 15% of the variation i BMD were associated with body weight, but only 8% with BMAD. Second, the Z scores compared with those for age-matched controls for BMD and BMC were -1.85 and -1.71, respectively; the Z score for BMAD was -1.64. Third, the precision error for BMC was reduced by expressing results as BMD (1.1 versus 0.5%); BMAD degraded precision slightly (0.7%). BMD appeared to be the optimal expression for bone densitometry because it provided the best diagnostic sensitivity and lowest precision error; there was a minimal influence of body size on BMD results. This study also showed that osteoporotic women, even in the first postmenopausal decade, had low spine BMD, small vertebral area, and low body weight. Such women may be particularly at risk of crush fracture.

Absorptiometry, Photon↗

Advances in noninvasive bone measurement.

Several noninvasive measurement methods are used for evaluation of metabolic disease. Single-photon (125I) scans of the peripheral skeleton are useful in some diseases but are ineffective in osteoporosis (even on the distal radius or os calcis) because they cannot predict spinal or femoral density. Also, peripheral measurements show high percentages of false negatives, that is many patients with fractures have normal peripheral density. Dual-photon (153Gd) scans of the spine, femur, and total skeleton are precise and accurate (2% error) and provide direct measurements of bone strength at fracture sites. This gives the best discrimination of abnormality and the most sensitive monitoring. Quantitative computed computed tomography (QCT) allows measurement of the spine but not the critical proximal femur area. QCT has a large accuracy error because (a) the limited area measured (under 5 cm3) fails to represent the total vertebral body, (b) technical errors, and (c) variable fat and osteoid influence the results.

Bone and Bones↗

Performance evaluation of a dual-energy x-ray bone densitometer.

We tested a dual-energy bone densitometer (LUNAR DPX) that uses a stable x-ray generator and a K-edge filter to achieve the two energy levels. A conventional scintillation detector in pulse-counting mode was used together with a gain stabilizer. The densitometer normally performs spine and femur scans in about 6 minutes and 3 minutes, respectively, with adequate spatial resolution (1.2 x 1.2 mm). Total body scans take either 10 minutes or 20 minutes. The long-term (6 months, n = 195) precision of repeat measurement on an 18-cm thick spine phantom was 0.6% at the medium speed. Precision error in vivo was about 0.6, 0.9 and 1.5% for spine scans (L2-L4) at slow, medium and fast speeds, while the error was 1.2 and 1.5 to 2.0%, respectively, for femur scans at slow and medium speed. The precision of total body bone density was 0.5% in vitro and in vivo. The response to increasing amounts of calcium hydroxyapatite was linear (r = 0.99). The densitometer accurately indicated (within 1%) the actual amount of hydroxyapatite after correction for physiological amounts of marrow fat. The measured area corresponded exactly (within 0.5%) to that of known annuli and to the radiographic area of spine phantoms. There was no significant effect of tissue thickness on mass, area, or areal density (BMD) between 10 and 24 cm of water. The BMD values for both spine and femur in vivo correlated highly (r = 0.98, SEE = .03 g/cm2) with those obtained using conventional 153Gd DPA. Similarly, total body BMD correlated highly (r = 0.96, SEE = .02 g/cm2) with DPA results.

Absorptiometry, Photon↗

Rapid determination of total hydroxyproline (HYP) in human urine by HPLC analysis of the phenylisothiocyonate (PITC)-derivative.

A recent development in the preparation for amino acid analysis is the use of phenylisothiocyanate (PITC) as a precolumn derivatizing agent prior to analysis to form a stable derivative. These derivatives can then be separated by reversed phase high pressure liquid chromatography (HPLC). Because of interest in the accurate measurement of urinary hydroxyproline (HYP), PITC was used for derivatization followed by HPLC analysis. Using an automated computer-controlled sampler system, up to 75 samples can be analyzed each 24 h. Technical details, specificity, and reliability of this method are provided. Duplicate measurements of HYP had a coefficient of variation (CV) of 5.5% and with a recovery of HYP in spiked urine samples of 94 to 104%. A sample frozen and thawed 10 times showed no change in the concentration of HYP. When total HYP was measured in 230 healthy women between 20 and 39 years of age, it was found to be 236.62 +/- 146.41 mumoles/L. These samples were researched in 3.5 days using this technique.

Adult↗

Bone density of the radius, spine, and proximal femur in osteoporosis.

Bone mineral density (BMD) was measured in 140 normal young women (aged 20 to 39 years) and in 423 consecutive women over age 40 referred for evaluation of osteoporosis. Lumbar spine and proximal femur BMD was measured using dual-photon absorptiometry (153Gd), whereas the radius shaft measurement used single-photon absorptiometry (125I). There were 324 older women with no fractures, of which 278 aged 60 to 80 years served as age-matched controls. There were 99 women with fractures including 32 with vertebral and 22 with hip fractures. Subsequently, another 25 women with hip fractures had BMD measured in another laboratory; their mean BMD was within 2% of that of the original series. The mean age in both the nonfracture and fracture groups was 70 +/- 5 years. The BMD in the age-matched controls was 20% to 25% below that of normal young women for the radius, spine, and femur, but the Ward's triangle region of the femur showed even greater loss (35%). The mean BMD at all sites in the crush fracture cases was about 10% to 15% below that of age-matched controls. Spinal abnormality was best discriminated by spine and femoral measurements (Z score about 0.9). In women with hip fractures, the BMD was 10% below that of age-matched controls for the radius and the spine, and the BMD for the femoral sites was about 25% to 30% below that of age-matched control (Z score about 1.6). Femoral densities gave the best discrimination of hip fracture cases and even reflected spinal osteopenia. In contrast, neither the spine nor the radius reflected the full extent of femoral osteopenia in hip fracture.

Adult↗

The relationship of peripheral to axial bone density.

Bone measurements were made in young normal women (n = 125) using single-photon absorptiometry (SPA with 125I) of the radius shaft and ultradistal radius, dual-photon absorptiometry (DPA with 153Gd) of the lumbar spine and the proximal femur, and 125I-based quantitative computed tomography (QCT) of the distal radius. Measurements of the appendicular and axial sites also were made in 21 women with spinal osteoporosis. The standard error of estimate in predicting axial densities from peripheral measurements was about 0.11 g/cm2 (10-13%) with all methods. The osteoporotic-normal difference was 21% for SPA of the radius shaft and 125I-QCT of the distal radius, 30% for the lumbar spine and 34% for the femoral neck. Prediction of spine and femur density, and diagnostic sensitivity, with 125I-QCT were equivalent to that obtained with conventional SPA, but the precision of 125I-QCT (0.8%) was 2-3 times better than both SPA and DPA.

Adult↗

The intragranular location of carboxyl groups in neuromelanin and lipofuscin in human brain and in meningeal melanosomes in mouse brain.

The intragranular location of carboxyl groups was tinctorially determined in human substantia nigra neuromelanin granules, human inferior olive lipofuscin granules, and mouse meningeal melanosomes. Soluble and insoluble lipid was stained with beta naphthol Sudans in unoxidized and oxidized frozen and paraffin sections containing neuromelanin or lipofuscin. Nile blue demonstrated carboxyls in unoxidized neuromelanin, lipofuscin, and melanin, and in oxidized neuromelanin and lipofuscin. Carbodiimide demonstrated carboxyls in unoxidized and oxidized lipofuscin and oxidized neuromelanin. In all instances, staining for carboxyls was inhibited by prior mild methylation, and proof of their presence was obtained by a pre-staining, stepwise, alternating, and repetitive mild demethylation, mild methylation sequence. Structurally, carboxyls were demonstrated in the neuromelanin granule's soluble lipid-free lipofuscin component, in the meningeal melanosome's melanin component, and virtually throughout the lipofuscin granule. The following structural and chemical basis was proposed for the different resistance of Nile blue staining of melanosomes and of neuromelanin and lipofuscin to acetone extraction. Nile blue forms an insoluble complex with melanosomal dopa-melanin's quinonoid, diphenolic, and undissociated carboxyl units. Such complex formation does not occur in neuromelanin's carboxyl-free dopamine-melanin component, however. Instead, Nile blue ionogenicly bonds with dissociated carboxyls belonging to the neuromelanin granule's lipofuscin component.

Aldehydes↗

The oxidative generation of sulfonic acid groups in neuromelanin and lipofuscin in the human brain.

Sulfonic acid groups were oxidatively generated in the soluble lipid-free lipofuscin component of neuromelanin of human substantia nigra and in lipofuscin of human inferior olive. Exposure of these oxidized, intraneuronal pigments to low pH Alcian blue or aldehyde fuchsin demonstrated an intensity of staining that related to the type of oxidant and the conditions of its use. Utilization of the following oxidants generated increasingly strong staining reactions as signified by the following sequence; periodic acid under mild conditions, bromine in carbon tetrachloride, hydrogen peroxide, periodic acid under drastic conditions, potassium permanganate followed by oxalic acid, hydrogen peroxide followed by bromine in carbon tetrachloride, potassium permanganate followed by metabisulfite or bisulfite, and performic acid. Neither Alcian blue nor aldehyde fuchsin revealed oxidatively generated aldehyde as judged by 1) their failure or near failure to stain inferior olive lipofuscin following mildly applied periodic acid, and 2) the increase in staining intensity, from moderate to strong, displayed by the soluble lipid-free lipofuscin component of neuromelanin and by inferior olive lipofuscin when potassium permanganate was followed by a rinse with metabisulfite or bisulfite in place of one with oxalic acid.

Brain↗

The dark brown integumentary pigment of a barnacle (Balanus eburneus). A comparative study.

A histochemical analysis involving tinctorial and solubility tests was pursued in conjunction with electron microscopy for the purpose of identifying the dark brown epidermal pigment of a barnacle (Balanus eburneus) as melanin and/or ommochrome. Histochemically, comparisons were made with other brown pigments located in the subcarapal epidermis of another crustacean, the fiddler crab (Uca pugilator), the dorsal skin of the red-backed salamander (Plethodon cinereus), the liver and testis of a slider turtle (Chrysemys sp.) and the substantia nigra of human brain. The solubility properties of the pigment of the two crustacean invertebrates were in general similar to one another and markedly different from the pigment of the three vertebrates. Insolubility in appropriate solvents classified the vertebrate pigment as melanin. The invertebrate pigment, however, which was soluble in the ommochrome solvents, concentrated sulfuric and formic acids and 2-chloroethanol, remained insoluble in the ommochrome solvents, dilute aqueous and methanolic hydrochloric acid and dilute sulfuric acid. On the basis of these solubilities, an unequivocal classification of the invertebrate pigment as either melanin or ommochrome did not appear possible. The tinctorial and electron microscopic properties of the barnacle pigment were also equally ambiguous in regard to its specific classification.

Animals↗

Histochemical observations on rodent brain melanin.

The presence or absence of melanin and neuromelanin in rodent brain was determined by means of light microscopic histochemistry. Melanin in melanocytes located in meninges and along cerebral capillaries occurred in abundance in C57 black mice and to a lesser extent in pigmented rats. Meningeal melanin bound cupric and ferrous ions, reduced ferricyanide, and appeared golden-brown in the bright field, light scattering in the dark field, and absorbant in the ultraviolet. Eleven aging albino rats were utilized to ascertain if the age related pigment, neuromelanin was present in neurons of the substantia nigra. Evidence of neuromelanin was not obtained as the cupric and ferrous ion uptake reactions were negative and neuromelanin could not be seen in the bright or dark field. Substantia nigra neurons did exhibit histochemical reactions for lipofuscin, however, as acid phosphatase positive, periodic acid-Schiff positive, and autofluorescent granules were demonstrated.

Aging↗

The presence of ethylenic bonds and vic-glycol groups in neuromelanin and lipofuscin in the human brain.

Through use of oxidation and blockading reactions, chemical group precursors of aldehyde demonstrable with Schiff reaction staining were identified in the soluble lipid-free lipofuscin component of neuromelanin of human substantia nigra and in lipofuscin of human inferior olive. Aldehyde generation was implied by moderate staining after bleaching neuromelanin and oxidizing lipofuscin with potassium permanganate followed by oxalic acid. Confirmation of aldehyde generation was achieved when diminished staining followed a sulfite addition blockade obtained by replacing oxalic acid with metabisulfite or bisulfite as well as by condensation blockades obtained with phenylhydrazine or aniline without replacing oxalic acid. Vic-glycol precursors of aldehyde were demonstrated in both pigments when acetylation or bromination preceded permanganate-oxalic acid and staining was unequivocally diminished only after acetylation. Vic-glycols were also demonstrated in lipofuscin by diminished staining when acetylation preceded periodic acid oxidation. Ethylenic precursors of aldehyde were suggested in performic acid-bleached neuromelanin when the minimal staining that followed this peracid's generation of Schiff reaction-negative dihydroxy groups became greatly intensified following an additional oxidation with periodic acid. This additional oxidation converted the dihydroxys to Schiff reaction-positive aldehyde. Ethylenes in lipofuscin were indicated when bromination before performic acid reduced subsequent staining.

Acetylation↗

Interference filter microfluorometry of neuromelanin and lipofuscin in human brain.

Following the use of a modified procedure for interference filter microfluorometry, bleached human substantia nigra neuromelanin exhibited a bimodal fluorescence maxima at 435 and 464 nm and human inferior olive lipofuscin, whether oxidized or not, exhibited a peak of fluorescence at 464 nm. Although the fluorescent component of bleached neuromelanin displayed its emission maxima in the spectral region characteristic for lipofuscin, and is regarded as such, its unique bimodality could represent some atypical, but as yet unknown, chemical property. Spectra of three fluorescence standards and a glass blank were also determined.

Aged↗

Acid fast staining of oxidized neuromelanin and lipofuscin in the human brain.

Acid fast staining of the bleached residuum of substantia nigra neuromelanin and of oxidized inferior olive lipofuscin was demonstrated in paraffin and frozen sections stained with the acetic acid, carbol fuchsin method of Barbeito-Lopez and the aldehyde fuchsin method of Gomori. Acid fast staining occurred when sections were exposed to a prestain oxidation with potassium permanganate in conjunction with a poststain differentiation with dilute hydrochloric acid. The acid fast staining with acetic acid, carbol fuchsin was differentiable and in contrast to the acid fast staining with aldehyde fuchsin which was nondifferentiable. A possible histochemical basis for differentiable and nondifferentiable acid fast staining was discussed. The identify of the bleached residuum of neuromelanin as lipofuscin was also discussed.

Adult↗