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Relationship between adenylate cyclase activity and regional myocardial energetics in experimental left ventricular hypertrophy.

The aim of this study was to examine the responsiveness of the hypertrophied left ventricle to beta-adrenergic stimulation in a pressure overload model produced by valvular aortic stenosis and characterized by reduced beta-adrenoceptor number. The study was designed to correlate changes in global and regional cardiac work and energetics in response to isoproterenol with adenylate cyclase activity. Eleven anesthetized dogs with left ventricular hypertrophy and 11 controls were studied at rest and during 0.5 and 1.0 micrograms/kg/min isoproterenol infusion. We measured regional work from segment length and force changes with ultrasonic dimension crystals and miniature force gauges in addition to arterial and left ventricular blood pressure and cardiac output. Regional myocardial oxygen consumption was calculated from O2 extraction using microspectrophotometry and blood flow using radioactively labeled microspheres. Adenylate cyclase activity was assayed at baseline and after stimulation with forskolin. Isoproterenol significantly increased heart rate, dP/dtmax, cardiac output, and external work to similar levels in control and hypertrophied animals. Similarly, regional work increased from 463 +/- 115 to 995 +/- 584 g x mm/min for controls and from 392 +/- 156 to 1175 +/- 577 for hypertrophied dogs with high dose isoproterenol. Regional O2 consumption also increased to similar levels (20.3 +/- 14.7 vs 16.2 +/- 6.3 ml O2/min/100 g) in both groups. Adenylate cyclase activity was lower in hypertrophy at baseline (23.9 +/- 7.3 vs 62.9 +/- 14.2 pM/min/mg protein for controls), but was the same as for controls with forskolin stimulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenylyl Cyclases↗

Relationship between cyclic-AMP content, regional myocardial function and O2 consumption in experimental left ventricular hypertrophy: effect of negative inotropes.

The aim of this study was to examine the hypothesis that negative inotropic agents that lower myocyte cyclic-AMP by different means would have similar effects on local myocardial segment work and O2 consumption in control hearts, but that this response would differ in left ventricular hypertrophy (LVH) induced by aortic valve stenosis. Open chest anesthesized LVH and control dogs were studied before and during esmolol (100 micrograms/kg/min) and acetylcholine (100 micrograms/kg/min) infusion. Regional work was calculated as the integrated product of instantaneous force (miniature transducer) and shortening (sonomicrometry) per min. Regional O2 consumption was calculated from blood flow (radioactive microspheres) and O2 saturation of small frozen vessels (microspectrophotometry). Cyclic-AMP level was determined with a competitive binding assay using 3H-cyclic-AMP and was found to be 731 +/- 90 (mean +/- S.D.) pmol/g in control and 711 +/- 163 in LVH. There were similar decreases in cyclic-AMP levels in control hearts with acetylcholine (365 +/- 135) and the beta adrenergic blocker (430 +/- 95). In LVH, esmolol lowered cyclic-AMP (383 +/- 39), but acetylcholine did not (689 +/- 105). In control animals, regional O2 consumption (7.7 +/- 0.6, 5.6 +/- 0.4 and 5.6 +/- 0.5 ml O2/min/100 g, control, acetylcholine, esmolol, respectively) and segment work (878 +/- 82, 546 +/- 80, 627 +/- 66 g*mm/min) fell to similar levels with these agents. Similar decreases were found in LVH with esmolol for O2 consumption (7.1 +/- 1.2, 5.1 +/- 1.0, baseline, esmolol) and segment work (895 +/- 140, 427 +/- 65). Acetylcholine had no significant effect on segment work (800 +/- 201), but did lower regional O2 consumption (4.0 +/- 0.7) in LVH dogs. It is concluded that there is a strong relationship between the level of cyclic-AMP and myocardial function and O2 consumption in control hearts. The action of acetylcholine is altered in LVH leading to an uncoupling between regional cyclic-AMP, function and metabolism.

Analysis of Variance↗

Cellular mechanisms for color-coding in holostean retinas and the evolution of color vision.

Electrophysiological recording and microspectrophotometry were used to analyze retinal function in representatives of the two surviving genera of holostean grade fish--the bowfin (Amia calva) and gars (Lepisosteus sp.). The properties of the cone photopigments, horizontal cells and ganglion cells show that these holostean retinas have cellular mechanisms for color vision which are fundamentally similar to those previously described for teleosts, turtle and mammals. These findings suggest that trichromatic receptor systems and opponent color-coding mechanisms may have evolved in primitive Neopterygii or more ancient fish, before the advent of teleosts. In conjunction with other recent data on living representatives of primitive fishes, these findings also add renewed plausibility for the view that vertebrate color vision could have taken a common origin some 400 million years ago from an ancestral aquatic jawed vertebrate.

Action Potentials↗

Color fundamentals deduced from carp ganglion cell responses.

Retinal ganglion cell responses were used to derive the action spectra (fundamentals) of the photopic color mechanisms in carp. Both silencing and chromatic adaptation techniques were used. The former proved to be more successful than the latter, because weak (even subthreshold) dynamic signals in one mechanism (mostly the red) caused strong effects on the other mechanisms. With chromatic adaptation these interacting signals were unavoidable, whereas with the silencing technique the modulation of the silenced mechanism was eliminated. The resultant pure action spectra agree well with the electrophysiological data of cone pedicles and horizontal cells. They differ, however, from the log-absorption specta found by microspectrophotometry. Apart from the differences in the slope of the long wavelength tails, the spectra also differ in the short wavelength region. The latter may indicate that in A-2 pigments, the beta-band absorption has a lower efficiency than the main-band absorption.

Action Potentials↗

Microspectrophotometric study of visual pigments in five species of geckos.

Photoreceptors and visual pigments were studied by light microscopy and microspectrophotometry in geckos Teratoscincus scincus, Gymnodactylus russovi, G. caspius, G. fedchenkovi and G. kotschui. Type A single, type B double and the thicker members of type C double cells were found to contain a green-sensitive visual pigment with the maximum absorption at 534-537 nm. Blue-absorbing pigment (lambda m = 446-460 nm) was only found in the thinner members of type C doubles.

Animals↗

Visual pigments and the labile scotopic visual system of fish.

Among mammals, birds, most reptiles and chondrichthians, only rhodopsins are present. Among agnathans, osteichthians, amphibians and certain freshwater turtles there are species having only porphyropsins or only rhodopsins or, more interestingly, both pigments, either sequentially or together. This latter grouping represents the paired-pigment species. Associated with the presence of paired-pigments is the possibility that the proportions of rhodopsin and porphyropsin may change. Depending on the characteristics of each paired-pigment species, naturally occurring changes in visual pigment ratios are related to migrations in anadromous and catadromous teleosts and anadromous cyclostomes and to seasonal variation in several teleosts. In addition, the visual pigment composition of certain species of teleosts has been altered by the specific effects of light, temperature, diet and hormones. Of two possible mechanisms for altering spectral sensitivity, varying the proportion of rhodopsin and porphyropsin is far more common than utilizing a single chromophore and changing the opsin. In addition to the long established evidence that extractable rod pigment ratios may change during the life cycle or in response to specific exogenous factors, there is the more recent recognition from microspectrophotometry that cone pigment ratios may also change in concert. The effect of lighting conditions and temperature on the visual pigment composition of certain paired-pigment species is presented.

Animals↗

The cone oil droplets of avian retinas.

The cone oil droplets of 19 species of birds from 11 families were examined by microspectrophotometry. Individual droplets were expanded with mineral oil, suspended in aqueous glycerol, and absorbance spectra measured between 700 and 320 nm. A classification of oil droplets is proposed, in which objective measurements of their carotenoids are related to the size, position and visual appearance of the droplets under the microscope. Some droplets contain no carotenoid and are transparent at wavelengths longer than 320 nm. Other droplets appear colorless but contain carotenoids absorbing at 385 or 402 nm. The pale droplets that have traditionally been described as greenish contain a mixture of two carotenoids. All of these types are distinct from yellow and red droplets. Red droplets contain astaxanthin esters, and yellow droplets contain a carotenoid with a spectrum similar to zeaxanthin. The 402 nm chromophore is galloxanthin, a C27 apo-carotenoid with 8 double bonds. The in vivo optical densities are 1-4 in the paler droplets, range up to about 8 in the yellow droplets, and can exceed 20 in the red droplets. All droplets that contain carotenoid can exhibit substantial absorption in the near u.v. The frequencies of the several droplet types in the retinas of different species suggests that these organelles respond readily to natural selection and may be involved in more than one function.

Animals↗

Larval and adult visual pigments of the zebrafish, Brachydanio rerio.

Photoreceptors of adult and larval zebrafish were assayed by microspectrophotometry to characterize their visual pigments, and to determine when the visual pigments first appear during development. Short single cone outer segments contained a pigment with a lambdamax near 417 nm. Long single cones and long members of double cone outer segments contained a pigment with a lambdamax near 480 nm. Short members of double cone outer segments contained a pigment with a lambdamax near 556 nm. Rod outer segments contained a rhodopsin with a lambdamax near 501 nm. All four visual pigment types found in adult photoreceptors were present in the earliest measurable larval photoreceptors.

Aging↗

"Self-screening" of rhodopsin in rod outer segments.

Microspectrophotometry (MSP) shows rhodopsin highly concentrated (about 3.0 mmol/l) in rod outer segments (ROS). Calculation of the in vivo absorption spectrum of human rhodopsin from such data reveals a striking failure to agree with the action spectrum of human rod vision. Agreement is good between the spectral distribution of absorption coefficients and the action spectrum, but the "concentration-broadening" (or "self-screening") introduced by the high end on absorbance at this concentration results in a misfit among the largest in the 93 years comparisons of this kind have been made! To deal with this anomaly, it has been suggested that "concentration-broadening" is inappropriate for rhodopsin in rod vision. This proposal was tested by comparing rod action spectra of 15-day-old and adult rats, since the lengths of ROS increase by a factor of about two in maturation. Three lines of evidence are inconsistent with it. Although the conundrum remains unexplained, it cannot be dismissed by supposing "self-screening" inappropriate for night vision.

Aging↗

A short-wavelength sensitive cone mechanism in juvenile yellow perch, Perca flavescens.

The retinal cone mosaic of adult yellow perch, Perca flavescens, consists of square units each having a central single cone surrounded by four identical twin cones. No cones are present at the corners of the square units. However, the cone types and organization of the retinal mosaic in juvenile P. flavescens has not been thoroughly described. We report here the presence of small, single cones at the corner positions of the square units in juvenile P. flavescens. Not every corner, nor every unit has these cones, and their distribution over the retina is not uniform. Microspectrophotometry places the visual pigment absorbance maximum of these short single cones in the region of 400 nm. Electrophysiological measurements of spectral sensitivity demonstrate a short-wavelength mechanism in juveniles that is absent, as are the small corner cones, in adults. Possible advantages of such a mechanism to juvenile perch are discussed.

Animals↗

The intrarhabdomal filters in the retinas of mantis shrimps.

The intrarhabdomal filters in the photoreceptors of compounds eyes of 32 species of mantis shrimps (Crustacea: Stomatopoda), representing seven families within the superfamilies Lysiosquilloidea and Gonodactyloidea, were surveyed by microspectrophotometry of filters in fresh cryosections of the retina. A total of up to four classes of filters exist in stomatopods: two each in Rows 2 and 3 of the midband. All lysiosquilloid species lacked the proximal filter in Row 3; a few also lacked the proximal Row 2 class. While most gonodactyloid species had all four possible classes, in some species the proximal filters of Row 3 and (in one case) Row 2 were missing. In all, at least 11 distinct spectral classes of pigments were found. Absorption spectra suggested that the filter pigments were probably carotenoids or carotenoproteins, although the distal filter of Row 3 was often exceptional, appearing to contain a mixture of pigments. While the types of pigments found in the filters of the various species generally followed taxonomic lines, numerous exceptions were found that were apparently related to the ecological requirements of the various species.

Animals↗

A method for the relative quantification of lipofuscin based on a computer image analysis system.

The accumulation of lipofuscin in neurons is the best recognized microscopic alteration of the aging brain. The broad spectrum which characterizes this progressive accumulation of pigment has made necessary the use of quantitative methods for its assessment. Present day techniques for the quantitation of lipofuscin (e.g. fluorescent microspectrophotometry and planimetry) are cumbersome, rely on manual techniques and may be limited by fluorescent artifacts. In this article we report a computer image analysis method for the relative quantification of lipofuscin in slides stained with a modified aldehyde fuchsin stain. The method preserves the topography of the pigment, is not limited by artifacts due to fluorescence, and has excellent inter-rater reliability.

Brain↗

Ribosomal RNA sequence conservation and gene number in the larval brine shrimp.

The haploid genome size of Artemia is determined to be about 0.9 X 10(12), as evidenced both by Feulgen microspectrophotometry of individual diploid class nuclei, which are but one of five polyploid classes present within the larvae, and by analysis of the reassociation kinetics of the isolated single copy DNA component. Polysomes isolated from 24-h incubation stage larvae contain an average of 10 ribosomes per messenger RNA molecule. Their rRNAs are found to have sedimentation coefficients of 18 S and 26 S, corresponding to molecular weights of 0.70 X 10(6) and 1.40 X 10(6), respectively, as determined by polyacrylamide electrophoresis and also by sucrose density centrifugation. Denaturation in glyoxal followed by agarose gel electrophoresis shows that unlike deuterostome rRNAs, Artemia 26 S rRNA contains a cryptic nick about midway in the molecule, which is not found in the 18 S molecule. Isolated rRNAs were labelled in vitro with 125I and hybridized with filter-immobilized DNA to saturation, which occurred at 0.051% for Xenopus, and at 0.074% for Artemia. From these results, it is calculated that in the haploid Artemia genome there are about 320 copies of the (18 S + 26 S) ribosomal RNA genes. Reciprocal heterologous hybridizations between these two species show that they share about 30% homology between their rDNA coding sequences.

Animals↗

Effect of exercise on age-related muscle atrophy.

Fifty-five-week-old male Wistar rats exercised voluntarily in running wheels for 10 weeks. The number and cross-sectional area of fibers in the slow-twitch soleus (SOL) and fast-twitch tibialis anterior (TA) muscles were recorded and compared with those of 20-week-old rats, and of age-matched rats not exercised. Muscle fibers were classified as fast-twitch oxidative glycolytic (FOG), fast-twitch glycolytic (FG), or slow-twitch oxidative (SO). After injection of horseradish peroxidase into the SOL and TA for retrograde neuronal labeling, oxidative enzyme activity of labeled motoneurons in the spinal cord was measured by microspectrophotometry. There were fewer FOG fibers in the SOL, and fewer FG fibers in the TA, at 65 weeks than at 20 weeks of age. The cross-sectional area of FOG and FG fibers in the TA was lower at 65 weeks than at 20 weeks of age. Exercise prevented the atrophy of FOG fibers in the TA. There were no age- or exercise-related differences in the number or oxidative enzyme activity of motoneurons in the SOL or the TA. These findings suggest that exercise can prevent the atrophy of FOG fibers by restoring their decreased metabolic capacity, and by inhibiting the degeneration of neuromuscular junctions.

Aging↗

Diagnostic significance of nuclear DNA content and nuclear area in oral hyperplasia, dysplasia, and carcinoma.

The nuclear DNA content and the nuclear area in hyperplasia, dysplasia, and carcinoma of the oral cavity were quantitatively analyzed by microspectrophotometry. In carcinoma, marked variability of the nuclear DNA content and the nuclear area was found. The mean nuclear DNA content, the variance of the nuclear DNA content, the frequency of cells above tetraploid (4 C), the mean nuclear area, and the variance of nuclear area in hyperplasia and dysplasia were significantly different from these values with carcinoma. In some cases of leukoplakia and verrucous hyperplasia with subsequent malignant formation, the values were beyond the lower limit of carcinoma. The results suggest that the nuclear DNA and the nuclear area are important objective indices for predicting malignant potential of precancerous lesions.

Carcinoma, Squamous Cell↗

DNA histochemical analysis in oral squamous cell carcinoma.

BACKGROUND: Recent years have witnessed an increasing emphasis on the role of nuclear DNA and its application in experimental pathological diagnosis to predict prognosis and management of certain neoplasm. AIMS: to establish objective criteria for the degree of differentiation and histochemical quantitative of nuclear DNA of oral squamous cell carcinoma (OSCC) using microspectrophotometric analysis. MATERIAL AND METHODS: The study was conducted on histologic materials from patient with OSCC. Two histological grading systems were used; Broder's and invasive front grading system were recorded. Microspectrophotometry was applied on Feulgen-stained sections to determine the quality of tumour nuclear DNA content in two different histological grading systems of OSCC. RESULTS: Nuclear DNA content increased significantly with decreasing tissue differentiation as well with increasing tumour size. CONCLUSION: The grading system and DNA content provides more objective and accurate criteria which relate the morphologic finding to biologic activity and growth patterns of oral cancer as compared to histologic differentiation alone.

Aneuploidy↗

Atrial natriuretic peptide reverses the negative functional effects of stunning in rabbit myocardium.

We tested the hypothesis that atrial natriuretic peptide (ANP) would decrease both the effects of myocardial stunning and oxygen consumption in rabbit hearts. In two groups of anesthetized open-chest rabbits, myocardial stunning was produced by two 15 min occlusions of the left anterior descending (LAD) artery separated by 15 min of reperfusion. Either ANP (0.2 mg) or vehicle (lactated Ringers) was then injected into the affected area of the left ventricle. In a third group, ANP was injected into the LAD region of non-stunned rabbits. Hemodynamic (heart rate, aortic and left ventricular pressures) and functional (wall thickening (WT), delay of onset of WT, and rate of WT) parameters were measured. Coronary blood flow (microspheres) and O2 extraction (microspectrophotometry) were used to determine myocardial O2 consumption. Stunning was demonstrated by an increase in the time delay to contraction and depressed WT. In the control group, baseline delay to contraction was 25+/-7 ms, and this increased to 84+/-16 following stunning and vehicle administration. In the ANP group, baseline delay was 20+/-6 at baseline and after stunning and ANP administration it was 30+/-7. Wall thickening decreased by approximately 30% with stunning and vehicle but only 8% in the ANP treated hearts. Stunning did not affect regional O2 consumption (6.0+/-1.1 stunned vs. 7.4+/-1.2 mlO2/min/100g non-stunned). ANP administration did not affect O2 consumption (7.3+/-1.7 stunned vs. 6.4+/-1.0 non-stunned). We therefore concluded that ANP administration reversed the effects of stunning without alteration in local O2 consumption in stunned myocardium.

Analysis of Variance↗

Recombination reaction of rhodopsin in situ studied by photoconversion of "indicator yellow".

We measured the kinetics of recombination of 11-cis-retinal with opsin in intact frog rod outer segment (ROS). The rhodopsin in ROS was bleached and allowed to decay to "indicator yellow," a photoproduct where all-trans-retinal is partly free, and partly bound to non-specific amino groups of disk membranes. By briefly illuminating the "indicator yellow" by an intense 465 or 380-nm flash, we then photoconverted all-trans-retinal to (mostly) the 11-cis- form thus introducing into ROS a certain amount of cis-chromophore. The recombination of cis-retinal with opsin and the formation of rhodopsin were followed by fast single-cell microspectrophotometry. Regeneration proceeded with a time constant of approximately 3.5 min; up to 27% of bleached visual pigment was restored. The regenerated pigment consisted of 91% rhodopsin (11-cis-chromophore) and 9% of presumably isorhodopsin (9-cis-chromophore). The recombination of 11-cis-retinal with opsin inside the ROS proceeds substantially faster than rhodopsin regeneration in the intact eye and, hence, is not the rate-limiting step in the visual cycle.

Animals↗