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PubMed1992

Expression of an oestrogen receptor-related antigen in human endometrium: a microphotometric study.

The expression of an oestrogen receptor-related antigen has been measured by microspectrophotometry in the glandular epithelium and stroma of tissue sections from endometrial biopsies of women administered exogenous replacement of sex steroids because of premature ovarian failure. We have demonstrated that previously dormant endometrium is capable of expressing an antigen related to oestrogen receptors and that its distribution is influenced by the sex steroid environment. Immunohistochemical staining was more intense (P less than 0.001) in the glandular compared to the stromal cells in tissue exposed solely to oestradiol. With the administration of progesterone, the stromal staining intensity increased (P less than 0.0001) and that of the glandular staining decreased (P less than 0.0001). No assumption, however, has been made to infer that colour is stoichiometrically related to the concentration of antigen present. Further validation of immunohistochemical procedures is required because histological localization and measurement of immunohistochemical staining may be straightforward, but its relationship to antigen concentration remains unproven.

Antibodies, Monoclonal↗
PubMed1996

Cerebral microregional oxygen balance during chronic versus acute hypertension in middle cerebral artery occluded rats.

This study was performed to compare microregional 0(2) supply and consumption balance in spontaneously hypertensive rats (SHR), normotensive Wistar Kyoto rats (WKY), and in phenylephrine-induced acutely hypertensive WKY (WKY + ph) rats. Under isoflurane anesthesia, a middle cerebral artery (MCA) of SHR (n = 7) and WKY (n = 14) rats was occluded. Seven of the WKY rats were infused with phenylephrine (WKY + ph) to keep the mean arterial pressure (MAP) at the same level as that of the SHR. In all animals, 1 h after MCA occlusion, regional cerebral blood flow (rCBF) was determined using an autoradiographic technique, and microregional arterial and venous 02 saturations were determined using microspectrophotometry. MAP was 76 +/- 4 (SD), 136 +/- 15, and 132 +/- 12 mm Hg for the WKY, WKY + ph, and SHR groups, respectively. All variables describing regional O2 balance and rCBF were similar between the SHR and the WKY groups in the ischemic cortex as well as in the contralateral cortex. With phenylephrine infusion, rCBF of both the ischemic cortex and the contralateral cortex were increased in the WKY group. The average 02 supply-to-consumption ratio in the ischemic cortex was higher in the WKY + ph than in the WKY or SHR group. In the ischemic cortex, heterogeneity of venous 02 saturation (SvO2), expressed as a coefficient of variation (CV = 100 X SD/mean), was significantly lower in the WKY + ph (18.3 +/- 2.4) group than in the SHR (30.5 +/- 11.8) or in the WKY (31.3 +/- 9.0) group. The number of veins with low 02 saturation (SvO2 < 40%) in the ischemic cortex was significantly lower in the WKY + ph than in the SHR or in the WKY group. Our data suggest that in chronically hypertensive animals, cerebrovascular adaptations enable the microregional 02 balance in focal ischemia to be maintained at a level similar to that of normotensive animals. However, in normotensive animals with focal cerebral ischemia, an acute increase of MAP improves microregional O2 balance.

Acute Disease↗
PubMed2005

Cone topography and spectral sensitivity in two potentially trichromatic marsupials, the quokka (Setonix brachyurus) and quenda (Isoodon obesulus).

The potential for trichromacy in mammals, thought to be unique to primates, was recently discovered in two Australian marsupials. Whether the presence of three cone types, sensitive to short- (SWS), medium- (MWS) and long- (LWS) wavelengths, occurs across all marsupials remains unknown. Here, we have investigated the presence, distribution and spectral sensitivity of cone types in two further species, the quokka (Setonix brachyurus) and quenda (Isoodon obesulus). Immunohistochemistry revealed that SWS cones in the quokka are concentrated in dorso-temporal retina, while in the quenda, two peaks were identified in naso-ventral and dorso-temporal retina. In both species, MWS/LWS cone spatial distributions matched those of retinal ganglion cells. Microspectrophotometry (MSP) confirmed that MWS and LWS cones are spectrally distinct, with mean wavelengths of maximum absorbance at 502 and 538 nm in the quokka, and at 509 and 551 nm, in the quenda. Although small SWS cone outer segments precluded MSP measurements, molecular analysis identified substitutions at key sites, accounting for a spectral shift from ultraviolet in the quenda to violet in the quokka. The presence of three cone types, along with previous findings in the fat-tailed dunnart and honey possum, suggests that three spectrally distinct cone types are a feature spanning the marsupials.

Animals↗
PubMed2005

Colour vision and speciation in Lake Victoria cichlids of the genus Pundamilia.

Lake Victoria cichlids are one of the most speciose groups of vertebrates. Selection on coloration is likely playing an important role in their rapid speciation. To test the hypothesis that sensory biases could explain species differences in mating preferences and nuptial coloration, we studied seven populations of four closely related species of the genus Pundamilia that differ in visual environment and male nuptial colour. Microspectrophotometry determined that the wavelength of maximum absorption (lambdamax) of the rod pigment and three cone pigments were similar in all four species. Only the long wavelength sensitive (LWS) pigment varied among species, with 3-4 nm shifts in lambdamax that correlated with differences in the LWS opsin sequence. These subtle shifts in lambdamax coincided with large shifts in male body colour, with red species having longer LWS pigments than blue species. Furthermore, we observed within and between species a correlation between water transparency and the proportion of red/red vs. red/green double cones. Individuals from turbid water had more red/red double cones than individuals from clear water. The variation in LWS lambdamax and in the proportion of red/red double cones could lead to differences in perceived brightness that may explain the evolution of variation in male coloration. However, other factors, such as chromophore shifts and higher order neural processing, should also be investigated to fully understand the physiological basis of differential responses to male mating hues in cichlid fish.

Africa, Eastern↗
PubMed1989

Comparative DNA analyses in longstanding ulcerative colitis with aneuploidy.

In seven patients with longstanding ulcerative colitis, multiple mucosal biopsies taken at colonoscopy were assessed for DNA content and histological dysplasia. DNA analyses were performed using flow cytometric analyses in biopsies and microspectrophotometry in imprint slides prepared from the biopsies. Abnormal aneuploid DNA content was detected in five of the patients in eight separate locations in the colon. There was good conformity between the two methods in the detection of aneuploidy. Both methods of DNA analysis are considered to be applicable in mucosal biopsies from patients with ulcerative colitis. In two patients with significant low grade dysplasia in association with macroscopical lesions (DALMs), there was a relationship with aneuploid DNA pattern found by flow cytometric or microspectrophotometric analyses. Indefinite changes, probably dysplastic were found in three other patients and two of those also displayed aneuploidy, although only once in the same biopsy location. In five locations aneuploidy was detected without concomitant dysplasia. DNA-aneuploidy seems therefore to appear earlier than dysplasia and may be an early marker of malignant transformation of the mucosa in ulcerative colitis. Prospective trials are needed before the clinical significance of early findings of aneuploidy can be determined.

Aneuploidy↗
PubMed1990

Microspectrophotometric DNA analysis in ulcerative colitis with special reference to its application in diagnosis of carcinoma and dysplasia.

The deoxyribonucleic (DNA) content was measured by microspectrophotometry in 100 specimens from 60 patients with ulcerative colitis, including six patients in whom the colitis was associated with carcinoma. Some 23 of 30 (77%) specimens of dysplastic tissue showed aneuploidy or polyploidy, whereas 50 of 53 (94%) specimens of non-dysplastic tissue showed diploidy. The difference was statistically significant (p less than 0.001). Polyploidy was often observed in non-dysplastic mucosa from patients who had carcinoma or dysplasia. In the non-dysplastic patients all samples of inflamed tissue showed diploidy. Some 10% of samples without inflammation, however, also showed polyploidy. A good correlation was found between the frequency of polyploid cells and the grade of dysplasia. Microspectrophotometric measurement of DNA content proved useful in the assessment and diagnosis of dysplasia in ulcerative colitis and could be considered for screening high risk patients.

Adult↗
PubMed1992

Image analysis of Feulgen-stained NIH/3T3 cells transformed with DNA of 4-nitroquinoline 1-oxide treated human breast epithelial cells.

The nuclear phenotypes of Feulgen-stained NIH/3T3 cells transformed with 4-nitroquinoline 1-oxide (4NQO) treated, human breast epithelial cell (HBEC) DNA were studied by scanning microspectrophotometry and image analysis and compared with data obtained for nontransformed cells and for NIH/3T3 cells under ras oncogene transfecting situations. The Feulgen-DNA content of the individual nuclei (NQ1, NQ2, and NQ3 phenotypes) of the transformed cells was found not to be deeply affected, although presence of chromatin structures resembling double minutes could be verified in part of the metaphases of the transformed cells. On the other hand, the chromatin supraorganization of these cells showed some changes involving increased (NQ2, NQ3) or decreased (NQ1) levels of condensation. The changes in chromatin packing states, however, were of small magnitude compared with those reported for NIH/3T3 cells transfected with a c-H-ras oncogene or an N-ras-containing MCF-7 cell DNA. It was assumed that the transformation of the NIH/3T3 cells is not always necessarily accompanied by high levels of chromatin condensation. The transformation of the NIH/3T3 cells induced by the 4NQO-treated HBEC DNA and particularly the changes in chromatin condensation in these transformed cells could not be attributed merely to a ras activation elicited by the carcinogen. It is suggested that a more complex transforming mechanism is involved, probably owing to the fact that a whole genomic DNA of the 4NQO-treated HBEC has been used for transfection.

3T3 Cells↗
PubMed2002

The retina of five atherinomorph teleosts: photoreceptors, patterns and spectral sensitivities.

We investigated the spectral and morphological features of the photoreceptors of five atherinomorph teleosts, representing two different orders, and with different life styles and habitats, the Beloniformes and Atheriniformes. The retinae of Belone belone (Belonidae), Dermogenys pusillus (Hemiramphidae), Atherina boyeri (Atherinidae), Marosatherina ladigesi (Telmatherinidae), and Melanotaenia maccullochi (Melanotaeniidae) were examined by light and electron microscopy and microspectrophotometry. In addition to rods, five morphologically different cone types were identified: short, intermediary and long single cones, and double cones which are arranged in distinct specific mosaics. Sporadically, triple cones were also found. Double cones were longer-wave-sensitive, but no general correlation between single cone morphology and spectral sensitivity could be demonstrated. The rods had lambda(max) close to 506-509 nm. The lambda(max) of cone visual pigments ranged from about 368 nm to 578 nm. Ultraviolet-sensitive single cones were present in the three freshwater species, M. ladigesi, M. maccullochi and D. pusillus and three spectrally distinct short-wave-sensitive single cone classes were identified in M. maccullochi. In M. ladigesi, spectral sensitivity varied among individuals due to varying rhodopsin/porphyropsin mixtures. In D. pusillus and M. maccullochi polymorphism of the longer-wave cone pigments might occur. These findings are discussed with respect to phylogeny, photic habitat, behavior and feeding habits.

Animals↗
PubMed1989

Fibronectin content of the annulus fibrosus in diabetic and non-diabetic sand rats.

Annulus fibrosus of intervertebral discs from diabetic and non-diabetic sand rats were examined by microspectrophotometry for fibronectin content. This was higher in the diabetic animals both in the dorsal and ventral parts and in the outer and inner lamellae of the annulus. It is suggested that diabetes-related changes in fibronectin are similar to changes in annular collagen observed in species other than sand rats.

Animals↗
PubMed1994

The DNA content of trophozoites and cysts of Giardia lamblia by microdensitometric quantitation of Feulgen staining and examination by laser scanning confocal microscopy.

We investigated direct measurement of the DNA content of the parasitic intestinal flagellate Giardia lamblia through quantitation by Feulgen microspectrophotometry and also by visualization of Feulgen-stained DNA chromosomes within dividing cells by laser scanning confocal microscopy. Individual trophozoites of Giardia (binucleate) contained 0.144 +/- 0.018 pg of DNA/cell or 0.072 pg DNA/nucleus. Giardia lamblia cysts (quadranucleate) contained 0.313 +/- 0.003 pg DNA or 0.078 pg DNA/nucleus. The genome size (C) value per nucleus ranged between 6.5-7.1 x 10(7) BP for trophozoites and cysts, respectively. Confocal microscopic examination of Giardia trophozoites undergoing binary fission revealed five chromosome-like bodies within each nucleus. Further information about genome size and DNA content within different Giardia species may help to clarify the pivotal role of these primitive eukaryotic cells in evolutionary development.

Animals↗
PubMed2003

Polymorphism of the rod visual pigment between allopatric populations of the sand goby (Pomatoschistus minutus): a microspectrophotometric study.

Absorbance spectra were measured by microspectrophotometry in retinal rods of sand gobies (Pomatoschistus minutus) from four allopatric populations (Baltic Sea, Swedish west coast, English Channel and Adriatic Sea). Mean (+/- S.E.M.) wavelengths of maximum absorbance (lambda(max)) were 508.3+/-0.5 nm, 505.4+/-0.2 nm, 506.2+/-0.3 nm and 503.0+/-0.3 nm, respectively. Pairwise comparison between the populations (post-ANOVA Scheffe's test) shows that each of the lambda(max) differences, except that between the Swedish west coast and the English Channel, is statistically significant (P<0.05). The shapes of the absorbance spectra indicated that the pigments were A1 rhodopsins with no measurable admixture of the A2 chromophore. Thus, the differences indicate polymorphism in the protein part (opsin) of the pigment. Convolution of A1 templates for lambda(max) values 508.3 nm and 503.0 nm with quantum spectra of the downwelling light at two locations at the south-west coast of Finland indicated that a 13-19% improvement in quantum catch would accrue in the Baltic environment from the 5.3 nm red-shift of the rod pigment of Baltic compared with Adriatic sand gobies.

Animals↗
PubMed2004

Interspecific and intraspecific views of color signals in the strawberry poison frog Dendrobates pumilio.

Poison frogs in the anuran family Dendrobatidae use bright colors on their bodies to advertise toxicity. The species Dendrobates pumilio Schmidt 1858, the strawberry poison frog, shows extreme polymorphism in color and pattern in Panama. It is known that females of D. pumilio preferentially choose mates of their own color morph. Nevertheless, potential predators must clearly see and recognize all color morphs if the aposematic signaling system is to function effectively. We examined the ability of conspecifics and a model predator to discriminate a diverse selection of D. pumilio colors from each other and from background colors. Microspectrophotometry of isolated rod and cone photoreceptors of D. pumilio revealed the presence of a trichromatic photopic visual system. A typical tetrachromatic bird system was used for the model predator. Reflectance spectra of frog and background colors were obtained, and discrimination among spectra in natural illuminants was mathematically modeled. The results revealed that both D. pumilio and the model predator discriminate most colors quite well, both from each other and from typical backgrounds, with the predator generally performing somewhat better than the conspecifics. Each color morph displayed at least one color signal that is highly visible against backgrounds to both visual systems. Our results indicate that the colors displayed by the various color morphs of D. pumilio are effective signals both to conspecifics and to a model predator.

Animals↗
PubMed2004

Multiple cone visual pigments and the potential for trichromatic colour vision in two species of elasmobranch.

Elasmobranchs (sharks, skates and rays) are the modern descendents of the first jawed vertebrates and, as apex predators, often occupy the highest trophic levels of aquatic (predominantly marine) ecosystems. However, despite their crucial role in the structure of marine communities, their importance both to commercial and to recreational fisheries, and the inherent interest in their role in vertebrate evolution, very little is known about their visual capabilities, especially with regard to whether or not they have the potential for colour vision. Using microspectrophotometry, we show that the retinae of the giant shovelnose ray (Rhinobatos typus) and the eastern shovelnose ray (Aptychotrema rostrata) contain three spectrally distinct cone visual pigments with wavelengths of maximum absorbance (lambdamax) at 477, 502 and 561 nm and at 459, 492 and 553 nm, respectively. The retinae of R. typus and A. rostrata also contain a single type of rod visual pigment with lambdamax at 504 and 498 nm, respectively. R. typus, living in the same estuarine waters as A. rostrata, were found to have identical visual pigments to R. typus inhabiting coral reef flats, despite a considerable difference in habitat spectral radiance. This is the first time that multiple cone visual pigments have been measured directly in an elasmobranch. The finding raises the possibility that some species are able to discriminate colour--a visual ability traditionally thought to be lacking in this vertebrate class--and it is evident that the visual ecology of elasmobranchs is far more complex than once thought.

Animals↗
PubMed2005

Adaptations to an extreme environment: retinal organisation and spectral properties of photoreceptors in Antarctic notothenioid fish.

The Notothenioid suborder of teleosts comprises a number of species that live below the sea ice of the Antarctic. The presence of 'antifreeze' glycoproteins in these fish as an adaptation to freezing temperature has been well documented but little is known about the adaptations of the visual system of these fish to a light environment in which both the quantity and spectral composition of downwelling sunlight has been reduced by passage through ice and snow. In this study, we show that the red/long-wave sensitive (LWS) opsin gene is not present in these fish but a UV-sensitive short-wave sensitive (SWS1) pigment is expressed along with blue-sensitive (SWS2) and green/middle-wave sensitive (Rh2) pigments. The identity and spectral location of maximal absorbance of the SWS1 and Rh2 pigments was confirmed by in vitro expression of the recombinant opsins followed by regeneration with 11-cis retinal. Only the SWS2 pigment showed interspecific variations in peak absorbance. Expression of the Rh2 opsin is localised to double cone receptors in both the central and peripheral retina, whereas SWS2 opsin expression is present only in the peripheral retina. SWS1 cones could not be identified by either microspectrophotometry or in situ hybridisation, presumably reflecting their low number and/or uneven distribution across the retina. A study of photoreceptor organisation in the retina of two species, the shallower dwelling Trematomus hansoni and the deeper dwelling Dissostichus mawsoni, identified a square mosaic in the former, and a row mosaic in the latter species; the row mosaic in Dissostichus mawsoni with less tightly packed cone photoreceptors allows for a higher rod photoreceptor density.

Adaptation, Biological↗
PubMed2002

Vision in the peafowl (Aves: Pavo cristatus).

The visual sense of the Indian blue-shouldered peafowl Pavo cristatus was investigated with respect to the spectral absorption characteristics of the retinal photoreceptors, the spectral transmittance of the ocular media and the topographic distribution of cells in the retinal ganglion cell layer. Microspectrophotometry revealed a single class of rod, four spectrally distinct types of single cone and a single class of double cone. In the case of the single cone types, which contained visual pigments with wavelengths of maximum absorbance (lambda(max)) at 424, 458, 505 and 567 nm, spectral filtering by the ocular media and the different cone oil droplets with which each visual pigment is associated gives predicted peak spectral sensitivities of 432, 477, 537 and 605 nm, respectively. Topographic analysis of retinal ganglion cell distribution revealed a large central area of increased cell density (at peak, 35,609 cells mm(-2)) with a poorly defined visual streak extending nasally. The peafowl has a calculated maximum spatial resolution (visual acuity) in the lateral visual field of 20.6 cycles degrees(-1). These properties of the peafowl eye are discussed with respect to its visual ecology and are compared with those of other closely related species.

Animals↗
PubMed1990

Standard specimens for stain calibration: application to Romanowsky-Giemsa staining.

Standardized specimens with reproducible staining properties were fabricated from extracts of biological objects (bovine liver, nucleoprotamine and defatted muscle). The standard specimens were stained with two formulations of the Romanowsky-Giemsa stain (RG), using the same azure B and eosin Y. One formulation used methanol and Sorensen's buffer and the other DMSO and Hepes buffer as solvents. The standard specimens were stained either in the composite stain or in the individual dyes dissolved in the same solvents and at the same concentration as the composite stain. Solution spectroscopy demonstrated different spectra for the two formulations with some wavelength regions varying by more than an order of magnitude. The RG spectra were also very different from those of the individual dyes dissolved at the RG concentration in the respective solvents. The stained standard specimens were analyzed by microspectrophotometry and were found to have spectra similar to those of cell smears. Furthermore, the standard specimens were shown to be a repeatable substrate for stain uptake. The transmitted light intensity from random fields of the same standardized specimen varied +/- 5%. When specimens were stained at the same time, the specimen-to-specimen variation depended on preparation conditions and the measurement wavelength, but was as good as +/- 5% for some conditions. The quantitative stain performance of both formulations was studied and compared. The standardized specimens provide a tool for the quantitative study of staining processes and specimen preparation procedures and for stain calibration.

Animals↗
PubMed2001

Is stagnating flow in former feeding arteries an indication of cerebral hypoperfusion after resection of arteriovenous malformations?

OBJECT: The authors' goal in this study was to challenge the proposed mechanism of the occlusive hyperemia theory, in which it is asserted that stagnating flow in the former feeding arteries of cerebral arteriovenous malformations (AVMs) leads to parenchymal hypoperfusion or ischemia, from which postoperative edema and hemorrhage originate. METHODS: Cortical oxygen saturation (SaO2) was measured in 52 patients by using microspectrophotometry in areas adjacent to AVMs before and after resection. The appearance of the former feeding arteries was categorized as normal (Group A); moderately stagnating (Group B); and excessively stagnating (Group C) on postoperative angiographic fast-film series. Patients and SaO2 values were pooled accordingly and compared using analysis of variance and Duncan tests (p < 0.05). Angiographic stagnation times in former feeding arteries were correlated in a linear regression/correlation analysis with SaO2 data (p < 0.05). All values are given as the mean +/- standard deviation. The average median postoperative SaO2 in Group C (15 patients) was significantly higher than in Groups B (17 patients) and A (20 patients) (Group C, 75.2 +/- 8.5; Group B, 67.5 +/- 10.8; Group A, 67.1 +/- 12 %SaO2), as was the average postoperative increase in SaO2 (Group C. 25.9 +/- 14.9; Group B, 14.6 +/- 14; Group A, 11.1 +/- 14 %SaO2). Angiographically confirmed stagnation times were also significantly longer in Group C than in Group B (Group C, 5.6 +/- 2.5; Group B, 1.3 +/- 0.6 seconds). A significant correlation/regression analysis showed a clear trend toward higher postoperative SaO2 levels with increasing stagnation time. CONCLUSIONS: Stagnating flow in former feeding arteries does not cause cortical ischemia, but its presence on angiographic studies is usually indicative of hyperperfusion in the surrounding brain tissue after AVM resection. In the context of the pathophysiology of AVMs extrapolations made from angiographically visible shunt flow to blood flow in the surrounding brain tissue must be regarded with caution.

Adolescent↗
PubMed1994

Evidence of sexual reproduction in the protozoan parasite Leishmania (Kinetoplastida: Trypanosomatidae).

Parasitic protozoa of the genus Leishmania (Kinetoplastida: Trypanosomatidae) are generally thought to multiply by binary fission; however, data from quantitative microspectrophotometry indicate that nuclear fusion or sexual reproduction takes place in the intracellular amastigote form. Among several different Leishmania species, the mean +/- SD nuclear DNA content of all promastigotes (extracellular form) and of some amastigotes (intracellular form) in macrophages was 0.098 +/- 0.032 relative units; in contrast, other amastigotes within the same macrophage had a mean +/- SD nuclear DNA content of 0.219 +/- 0.050. The latter population of amastigotes are apparently produced when the nuclei of a pair of 0.098 amastigotes fuse. These 0.219 amastigotes later go through what is probably the typical meiotic cycle to reform the 0.098 condition we observed among promastigotes. The demonstration of sexual reproduction in Leishmania has important implications for the future direction of research on this medically important parasite.

Animals↗