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Microscopic evaluation of the sequential degeneration and repair of the retinae of albino mice after high-dose laser irradiation.

After irradiation of the retinae of albino mice with a dose of 13 J, using a ruby pulse laser, the lesions were followed for a period of 60 days. Severe degenerations were observed in the outer layer 2-9 days after irradiation. By the 21st day after irradiation, the pigment epithelium started to regenerate and outer segments began to reform. Progressive recovery and repair were by and large completed by the 60th day after irradiation.

Animals↗

Reinnervation by axon collaterals from single facial motoneurons to multiple muscle targets following axotomy in the adult guinea pig.

To study the process of recovery from facial palsy experimentally, the location of cranial motoneurons supplying the posterior belly of the digastric muscle (PDG) and the extratemporal portion of the facial nerve trunk was examined in a double-labeling paradigm using two retrograde tracers in the adult guinea pig of which the facial nerve had been surgically injured. In different stages after the induced facial palsy had recovered functionally (4-13 weeks after the surgical operation), wheat germ-agglutinated horseradish peroxidase (WGA-HRP) was injected into the PDG and Fluoro-Ruby (FR) was applied to the proximal cut end of the extratemporal portion of the facial nerve trunk. Distribution of neurons retrogradely labeled with WGA-HRP and/or FR was plotted in the brainstem and compared with that of the controls. In the intact cases, HRP-labeled neurons were restrictedly seen in the accessory facial nucleus (Acs7), while FR-labeled neurons were found within the main facial nucleus (FMN). In the axotomized cases: (1) HRP-labeled neurons were seen diffusely in the Acs7 as well as in the FMN, where normal myotopical representation no longer seemed to be maintained. (2) FR-labeled neurons were also observed diffusely in the FMN and the Acs7. (3) A considerable number of neurons were doubly labeled with WGA-HRP and FR in both the Acs and the FMN in cases with shorter survival periods (4-7 weeks), but not in cases with longer survival periods (12-13 weeks). Thus, new findings show that connections are temporarily maintained by single, facial motoneurons with axon collaterals to multiple muscle targets in adult mammals.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Urokinase-treatment of fresh laser irradiation-induced vitreous hemorrhage.

Using a ruby laser, we induced vitreous hemorrhage in each eye of 12 rabbits. On the following day either 25,000 Ploug units of urokinase or saline were injected into the eyes of each rabbit. Urokinase did not accelerate the absorption of blood from the vitreous but prevented the development of severe vitreous fibrosis which replaced the hemorrhage in seven of the control eyes.

Animals↗

The Minnesota Grading System of eye bank eyes for age-related macular degeneration.

PURPOSE: The Minnesota Grading System (MGS) is a method to evaluate human eye bank eyes and determine the level of age-related macular degeneration (AMD), by using criteria and definitions from the Age-Related Eye Disease Study (AREDS). METHODS: Donor eyes (108 pairs) from the Minnesota Lions Eye Bank were cut circumferentially at the pars plana to remove the anterior segment. A 1000 +/- 2.5-microm ruby sphere was placed on the optic nerve as a size reference. A digital, high-resolution, color macular photograph was taken through a dissecting microscope. The neurosensory retina was removed from one globe of the pair. The underlying retinal pigment epithelium was rephotographed, localizing the fovea with a proportional triangle. A grid was superimposed in the macular photographs and images were graded according to AREDS criteria. Twenty pairs were dissected bilaterally and graded for symmetry. RESULTS: Eighty-eight globes were graded into one of four MGS categories. Nineteen (95%) of 20 globes had symmetric grades. CONCLUSIONS: The MGS provides a methodology to grade donor tissue from eye bank eyes to correspond to the AREDS classification system. Donor tissue may be used for subsequent molecular analysis, including genomics and proteomics.

Aged↗

Superoxide production by phagocytic leukocytes: the scientific legacy of Bernard Babior.

It was 32 years ago that Bernard Babior, Ruby Kipnes, and I submitted a paper to the JCI reporting that polymorphonuclear leukocytes produce superoxide (O(2) (-)) during phagocytosis and that this highly reactive oxygen radical might function as a microbicidal agent. The story of how our lab came to this discovery is one of a special relationship between a student and his brilliant mentor.

History, 20th Century↗

Adipose energy stores, physical work, and the metabolic syndrome: lessons from hummingbirds.

Hummingbirds and other nectar-feeding, migratory birds possess unusual adaptive traits that offer important lessons concerning obesity, diabetes and the metabolic syndrome. Hummingbirds consume a high sugar diet and have fasting glucose levels that would be severely hyperglycemic in humans, yet these nectar-fed birds recover most glucose that is filtered into the urine. Hummingbirds accumulate over 40% body fat shortly before migrations in the spring and autumn. Despite hyperglycemia and seasonally elevated body fat, the birds are not known to become diabetic in the sense of developing polyuria (glucosuria), polydipsia and polyphagia. The tiny (3-4 g) Ruby-throated hummingbird has among the highest mass-specific metabolic rates known, and loses most of its stored fat in 20 h by flying up to 600 miles across the Gulf of Mexico. During the breeding season, it becomes lean and maintains an extremely accurate energy balance. In addition, hummingbirds can quickly enter torpor and reduce resting metabolic rates by 10-fold. Thus, hummingbirds are wonderful examples of the adaptive nature of fat tissue, and may offer lessons concerning prevention of metabolic syndrome in humans.

Adipose Tissue↗

Two-dimensional gel proteome reference map of blood monocytes.

BACKGROUND: Blood monocytes play a central role in regulating host inflammatory processes through chemotaxis, phagocytosis, and cytokine production. However, the molecular details underlying these diverse functions are not completely understood. Understanding the proteomes of blood monocytes will provide new insights into their biological role in health and diseases. RESULTS: In this study, monocytes were isolated from five healthy donors. Whole monocyte lysates from each donor were then analyzed by 2D gel electrophoresis, and proteins were detected using Sypro Ruby fluorescence and then examined for phosphoproteomes using ProQ phospho-protein fluorescence dye. Between 1525 and 1769 protein spots on each 2D gel were matched, analyzed, and quantified. Abundant protein spots were then subjected to analysis by mass spectrometry. This report describes the protein identities of 231 monocyte protein spots, which represent 164 distinct proteins and their respective isoforms or subunits. Some of these proteins had not been previously characterized at the protein level in monocytes. Among the 231 protein spots, 19 proteins revealed distinct modification by protein phosphorylation. CONCLUSION: The results of this study offer the most detailed monocyte proteomic database to date and provide new perspectives into the study of monocyte biology.

Journal Article↗

My life in testing.

This autobiographical essay describes my career as a psychodiagnostician, which began at the City College of New York in 1941 and ended in the late 1970s when I became a full-time psychoanalyst in Manhattan. As a green, 20-year-old psychology undergraduate, I was picked by Gardner Murphy to assist David Rapaport at the Menninger Clinic in Topeka, Kansas, who was developing new methods for using psychological tests to diagnose and treat soldiers during World War II. Our findings were presented in the classic, two volume work, Diagnostic Psychological Testing, published in 1945-1946 (Rapaport, Gill, & Schafer, 1945-1946; 1968), to which I added two clinically enriched monographs: The Clinical Application of Psychological Tests, published in 1948 (Schafer, 1948/1995), and Psychoanalytic Interpretation in Rorschach Testing (Schafer, 1954), published in 1954. After the Menninger Clinic, I continued to hone my assessment expertise at the Austen Riggs Center and Yale University, but I also sought opportunities to develop psychotherapy skills. I completed psychoanalytic training in 1960 at the Western New England Institute for Psychoanalysis and thereafter sought positions that would offer more time for treating patients. A notable highpoint in my testing career was assessing Jack Ruby, killer of Lee Harvey Oswald. I look back fondly at my years as an assessment psychologist, which produced nearly 4 decades of great memories.

Forensic Psychiatry↗

Limits to flight energetics of hummingbirds hovering in hypodense and hypoxic gas mixtures.

Hovering hummingbirds offer a model locomotor system for which analyses of both metabolism and flight mechanics are experimentally tractable. Because hummingbirds exhibit the highest mass-specific metabolic rates among vertebrates, maximum performance of hovering flight represents the upper limit of aerobic locomotion in vertebrates. This study evaluates the potential constraints of flight mechanics and oxygen availability on maximum flight performance. Hummingbird flight performance was manipulated non-invasively using air and gas mixtures which influenced metabolism via variable oxygen partial pressure and/or altered flight mechanics via variable air densities. Limits to the locomotor capacity of hovering ruby-throated hummingbirds (Archilochus colubris) were unequivocally indicated by aerodynamic failure in either air/helium or air/heliox mixtures. Air/helium mixtures are hypodense and hypoxic; failure to sustain hovering flight occurred at 63% of the density of sea-level air and at an oxygen concentration of 12%. Air/heliox mixtures are hypodense but normoxic; failure in hovering occurred at 47% of sea-level air density. Thus, hummingbirds demonstrated considerable power reserves in hovering flight as well as hypoxic tolerance. In air/helium mixtures, hovering was limited by oxygen supply and not by flight mechanics. Birds hovering in air/helium mixtures increased their mechanical power output but not their rate of oxygen consumption. By contrast, birds hovering in air/heliox mixtures increased both mechanical performance and metabolic expenditure. Under hypoxia, hovering hummingbirds demonstrated non-negligible, but still limited, capacities for anaerobic metabolism and/or oxygen storage. Depending on the physical context, hummingbird flight performance can therefore be limited by oxygen availability or by flight aerodynamics.

Air↗

Hummingbird hovering performance in hyperoxic heliox: effects of body mass and sex.

Owing to their small size and hovering locomotion, hummingbirds are the most aerobically active vertebrate endotherms. Can hyperoxia enhance the flight performance of this highly oxygen-dependent group? Hovering performance of ruby-throated hummingbirds (Archilochus colubris) was manipulated non-invasively using hyperoxic but hypodense gas mixtures of sea-level air combined with heliox containing 35% O2. This manipulation sheds light on the interplay among metabolic power input, mechanical power output and aerodynamic force production in limiting flight performance. No significant differences in flight mechanics and oxygen consumption were identified between hyperoxic and normoxic conditions. Thus, at least in the present experimental context, hyperoxia did not change the major metabolic and mechanical parameters; O2 diffusive capacities of the respiratory system were probably not limiting to a significant extent. Compared with hummingbirds in our previous studies, the present experimental birds were heavier, had resultant shorter hover-feeding durations and experienced aerodynamic failure at higher air densities. Because hummingbirds have relatively stable wingbeat frequencies, modulation of power output was attained primarily through variation in stroke amplitude up to near 180 degrees. This result indicates that maximum hovering performance was constrained geometrically and that heavier birds with greater fat loads had less margin for enhancement of power production. Sexual dimorphism in flight adaptation also played a role, with males showing more limited hovering capacities, presumably as a trade-off for increased maneuverability.

Adipose Tissue↗

Hummingbird hovering energetics during moult of primary flight feathers.

How does a hovering hummingbird compensate for the loss of flight feathers during moult when the mechanism of lift force generation by flapping wings is impaired? The flight performance of five individual ruby-throated hummingbirds with moulting primary flight feathers and reduced wing area was compared with that before their moult. Hummingbirds were flown in reduced air densities using normoxic heliox so that a range of flight energetics was displayed. The rate of moulting and the extent of wing area loss varied among individuals. One female could tolerate a 30% loss of wing area in moulting and flew with only three outer primaries per wing. Further exploratory study using the artificial reduction of wing area, either by cutting the tips of the outer primaries of a male or by plucking the secondaries of two females, suggested that secondaries play a minor role in lift force generation during hovering whereas the tip area of primaries is crucial. For the five birds, ranges of whole-bird oxygen consumption rates, wingbeat kinematics (stroke amplitude) and lift coefficients did not vary during the moult. This constancy was mainly achieved through weight loss that alleviated aerodynamic force requirements for weight support during hovering. Since the metabolic power expenditure during moult was similar to that of normal birds but the mechanical power requirement was reduced, the flight efficiency also showed a sharp reduction during moult. This increased cost of flight may result from disruption of the integrity of the flight machinery. Overall, the control of body mass in hummingbirds can provide similar aerodynamic, muscle mechanical and physiological capacities under conditions of variable flight demand.

Animals↗

Hovering performance of hummingbirds in hyperoxic gas mixtures.

Hummingbirds evolved during a period of decline in atmospheric oxygen concentration and currently encounter varying levels of oxygen availability along their elevational distribution. We tested the hypothesis that inspiration of hyperoxic gas increases hummingbird hovering performance when birds are simultaneously challenged aerodynamically. We measured the maximum duration of hovering flight while simultaneously monitoring the rate of oxygen consumption of ruby-throated hummingbirds (Archilochus colubris) in low-density heliox that was either normoxic (21% O2) or hyperoxic (35% O2). As air density decreased below 0.85 kg x m(-3), hummingbirds hovered significantly longer in hyperoxia than in normoxia, but the air density at which the birds could no longer sustain hovering flight was independent of oxygen concentration. At low air densities in hyperoxia flight trials, hummingbirds appeared to increase their rate of oxygen consumption relative to flight sequences at equivalent densities in normoxia trials, but these differences were not significant. We tested the hypothesis that hummingbirds can discriminate between environments that differ in oxygen concentration. In another density-reduction experiment, hummingbirds were allowed to choose between artificial feeders infused with either normoxic or hyperoxic gases. The hypothesis was not supported because birds failed to associate oxygen concentration with a particular feeder independently of air density. Supplemental oxygen thus yields increased hovering duration at intermediate air densities, but the minimum density at which birds can fly is limited exclusively by aerodynamic considerations.

Air↗

Effect of SA1, a herbal formulation, on sexual behavior and penile erection.

SA1 is a mixture of 9 Oriental herbs (Korean red ginseng, fermented soybean, Tribulus terrestris, Fructus Rubi, Fructus Lycii, Semen Cuscutae, Dioscorea Rhizome, Fructus Corni and Fructus Crataegi) that are widely used as energizers and vitalizers in the indigenous system of medicine and have been alleged to improve the sexual functions in men. This study evaluated SA1 using both in vitro and in vivo experiments on laboratory animals in order to determine its effect on the sexual behavior and penile erection. The male rats used to examine the copulatory behavior were administered either the vehicle or SA1 (30, 100, 300, 600 mg/kg) orally for 2 weeks. The intracavernous pressure and systemic blood pressure were recorded in anesthetized rats. The responses to acetylcholine and SA1 of rabbit corpus cavernosum strips were also examined. There was an overall increase in the copulatory behavior parameters in the SA1-treated rats, which was reflected by a decrease in the mount and intromission latencies and an increase in the ejaculation latency and mount frequency. SA1 significantly increased the ratio of the intracavernous pressure to mean arterial pressure. In vitro, SA1 significantly enhanced the relaxation responses to acetylcholine. These results suggest that SA1 improves the sexual activity and erectile function.

Animals↗

Lasers in periodontology.

Since the development of the ruby laser by Maiman in 1960, lasers have been widely employed in medicine for a number of years. The purpose of this paper is to summarize potential applications for lasers in dentistry, with special regard to periodontology. This article briefly describes clinical applications of lasers and laser safety. Particularly, the use of a diode laser seems to be promising, especially in already compromised transplant patients, who need to be treated with a technique where the operative and post-operative blood loss, post-operative discomfort and the recurrence of drug-induced gingival overgrowth need to be kept to a minimum or eliminated. Therefore, the use of lasers in periodontology may lead to an alteration in present clinical practice and help to establish the best management strategy because, by maintaining periodontal health, the life quality of patients can be improved.

Dental Calculus↗

High-temperature and high-pressure cubic zirconia anvil cell for Raman spectroscopy.

A simple and inexpensive cubic zirconia anvil cell has been developed for the performance of in situ Raman spectroscopy up to the conditions of 500 degrees C and 30 kbar pressure. The design and construction of this cell are fully described, as well as its applications for Raman spectroscopy. Molybdenum heater wires wrapped around ceramic tubes encircling two cubic zirconia anvils are used to heat samples, and the temperatures are measured and controlled by a Pt-PtRh thermocouple adhered near the sample chamber and an intelligent digital control apparatus. With this cell, Raman spectroscopic measurements have been satisfactorily performed on water at 6000 bar pressure to 455 degrees C and on ice of room temperature to 24 kbar, in which the determinations of pressures make use of changes of the A1 Raman modes of quartz and the shift of the sharpline (R-line) luminescence of ruby, respectively.

Equipment Design↗

Factors determining the stability, resolution, and precision of a conventional Raman spectrometer.

We verified the performance of a conventional Raman spectrometer, which is composed of a 30 cm single polychromator, a Si based charge-coupled device (CCD) camera, and a holographic supernotch filter. For that purpose, the time change of the peak positions of Raman spectra of naphthalene and fluorescence spectra of ruby (Cr-doped Al(2)O(3)) were monitored continually. A time-dependent deviation composed of two components was observed: a monotonous drift up to 0.4 cm(-1) and a periodic oscillation with a range of 0.15 cm(-1). The former component was stabilized at approximately 2000 s after the CCD detector was cooled, indicating that incomplete refrigeration of the CCD detector induced the drift. The latter component synchronized with the periodic oscillation of the room temperature, indicating that thermal expansion or contraction of the whole apparatus induced this oscillation. The implemental deviation is reduced when measurements are conducted using a sufficiently cooled CCD detector at a constant room temperature. Moreover, the effect of the room temperature oscillation is lowered in a spectrum acquired over a duration that is longer than one cycle of this oscillation. Applying the least squares fitting method to carefully measured spectra enhanced the precision of the determination of the peak position to 0.05 cm(-1) using the spectrometer with pixel resolution of 1.5 cm(-1).

Journal Article↗

Visible and fluorescent staining of two-dimensional gels.

Staining of two-dimensional gel constitutes a crucial step in comparative proteome analysis with respect to the number of proteins analyzed, the accuracy of spot quantification, and compatibility with mass spectrometry. The present chapter describes procedures for several visible and fluorescent dyes compatible with mass spectrometry: colloidal Coomassie blue, silver nitrate, Sypro Ruby, Deep Purple, and 5-hexadecanoylamino-fluorescein.

Arabidopsis↗