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Detection of fluorescence dye-labeled proteins in 2-D gels using an Arthur 1442 Multiwavelength Fluoroimager.

Labeling of proteins with SYPRO Orange, SYPRO Red, and SYPRO Ruby after 2-D polyacrylamide gel electrophoresis (PAGE) using plastic-backed immobilized pH gradient (IPG) strips and precast SDS polyacrylamide gels was tested. Protein spots were detected using an Arthur 1442 Multiwavelength Fluoroimager. The labeling methods described allow detection of proteins both after isoelectric focusing (IEF) and PAGE with a sensitivity higher than or comparable to standard silver staining methods. In addition to the post-labeling methods mentioned above, pre-labeling with the cysteine-specific fluorophore monobromobimane before 2-D PAGE is a sensitive, fast, and cost-effective alternative to existing staining protocols.

Bacterial Proteins↗

Biomechanics of rugby union scrummaging. Technical and safety issues.

In the game of ruby union, the scrum epitomises the physical nature of the game. It is both a powerful offensive skill, affording a base for attacking play, and a defensive skill in denying the opposition clean possession. However, the scrum has also been implicated in a large proportion of serious spinal injuries in rugby union. The majority of injuries are found to occur at engagement where the forces experienced by front-row players (more than two-thirds of a tonne shared across the front-row) can exceed the structural limits of the cervical spine. These large forces are a consequence of the speed of engagement and the weight (and number) of players involved in the scrum. This highlights not only the need for physical preparation of all forwards but particularly player restraint at engagement, and justifies the 'crouch-pause-engage' sequence recently introduced to 'depower' the scrum. As the hooker is the player exposed to the greatest loads throughout the scrum and subsequently most at risk, he should determine the timing of engagement of the 2 front-rows. Stability of the scrum is an indication of front-row players' ability to utilise their strength to transmit the force to their opponents as well as the push of second-row and back-row players behind them in the scrum. This appears to be independent of the size of players. Equally, it reflects the risk of chronic degeneration of the musculoskeletal system through repeated exposure to these large stresses. However, not only are older and more experienced players better able to generate and transmit these forces, they are also able to maintain the integrity of the scrum. A large proportion of individual players' efforts to generate force is lost in their coordinated effort in a normal scrum. It is assumed these forces are dissipated through players re-orientating their bodies in the scrum situation as well as to less efficient shear forces and to the elastic and compressive tissues in the body. It again reinforces the importance of physical preparation for all forwards to better withstand the large forces involved in scrummaging. Despite negative publicity surrounding the risk of serious spinal injury in rugby union, limited research has been conducted to examine either the mechanisms of injury or techniques implicated in causing injury. Biomechanical information can provide systematic bases for modifying existing techniques and assessing the physical capacities necessary to efficiently and safely play in the serum. This will both improve performance of game skills and minimise the potential for injury.

Biomechanical Phenomena↗

Laser hair removal: guidelines for management.

Laser-assisted hair removal is the most efficient method of long-term hair removal currently available. Several hair removal systems have been shown to be effective in this setting: ruby laser (694nm), alexandrite laser (755nm), diode laser (800nm), intense pulsed light source (590 to 1200nm) and the neodymium:yttrium-aluminium-garnet (Nd:YAG) laser (1064nm), with or without the application of carbon suspension. The parameters used with each laser system vary considerably. All these lasers work on the principle of selective photothermolysis, with the melanin in the hair follicles as the chromophobe. Regardless of the type of laser used multiple treatments are necessary to achieve satisfactory results. Hair clearance, after repeated treatments, of 30 to 50% is generally reported 6 months after the last treatment. Patients with dark colored skin (Fitzpatrick IV and V) can be treated effectively with comparable morbidity to those with lighter colored skin. Although there is no obvious advantage of one laser system over another in terms of treatment outcome (except the Nd:YAG laser, which is found to be less efficacious, but more suited to patients with darker colored skin), laser parameters may be important when choosing the ideal laser for a patient. Adverse effects reported after laser-assisted hair removal include erythema and perifollicular edema, which are common, and crusting and vesiculation of treatment site, hypopigmentation and hyperpigmentation (depending on skin color and other factors). Most complications are generally temporary. The occurrence of hypopigmentation after laser irradiation is thought to be related to the suppression of melanogenesis in the epidermis (which is reversible), rather than the destruction of melanocytes. Methods to reduce the incidence of adverse effects include lightening of the skin and sun avoidance prior to laser treatment, cooling of the skin during treatment, and sun avoidance and protection after treatment. Proper patient selection and tailoring of the fluence used to the patient's skin type remain the most important factors in efficacious and well tolerated laser treatment. While it is generally believed that hair follicles are more responsive to treatment while they are in the growing (anagen) phase, conflicting results have also been reported. There is also no consensus on the most favorable treatment sites.

Clinical Protocols↗

Holographic investigation of the impact response of human heads.

The impact response of intact human cadaver heads was studied by means of double-pulsed holography using a ruby laser, and the transient deformation process at the surface of the skull was recorded. Frequency and acceleration measurements following impact were obtained. On frontal impact, the shock wave reached the opposite pole after 0.2 to 0.3 msec, and maximum amplitudes were found in the temporal regions. On impact to the vault, the deformation wave reached the skull base after 0.15 to 0.175 msec. Centers of maximum amplitude were recorded in the parietal and temporal regions. Deviations in the interference lines occurred at some sutures, revealing discontinuities. High-frequency components of the shock wave were demonstrated. The authors believe that fracture mechanisms may be elucidated by further studies of impact responses.

Craniocerebral Trauma↗

Shellac formulations to reduce epiphytic survival of coliform bacteria on citrus fruit postharvest.

Survival of the coliform bacteria Enterobacter aerogenes and Escherichia coli was monitored in a neutral carboxymethylcellulose formulation and in shellac formulations with various pH and concentrations of ethanol and the preservative paraben; populations were subsequently measured from the surface of citrus fruit coated with these formulations. Numbers of the two bacteria increased over 24 h from 10(6) CFU/ml to approximately 10(8) CFU/ml in the carboxymethylcellulose solution, but over this time numbers remained little changed in the neutral solution of shellac. The Enterobacter was more tolerant of alcohol over a 3-h period: although its numbers in a shellac solution with 10% ethanol dropped from more than 10(6) CFU/ml to just over 10(3) CFU/ml. E. coli and a third species. Klebsiella pneunoniae, declined toward the limit of detection (5 CFU/ ml) during this time. The addition of morpholine to increase the formulation pH to 9.0 caused numbers of bacteria to plummet to an undetectable level within 30 to 60 min. On Ruby Red grapefruit and Valencia oranges in storage at 13 degrees C numbers of E. aerogenes and E. coli declined over 2 weeks from 10(5) CFU/cm2 to less than 2.5 x 10(1), but most of the loss in numbers occurred within 1 day. Numbers remained significantly less on shellacked fruit compared with those applied in the carboxymethylcellulose coating, and a shellac coating prepared from a pH 9 solution was more toxic to these species than one in which 12% ethanol had been added to the neutral formulation. The addition of the preservative paraben in the basic shellac was further inhibitory.

Carboxymethylcellulose Sodium↗

Residues of the quinone outside inhibitor fungicide trifloxystrobin after postharvest dip treatments to control Penicillium spp. on citrus fruit.

The effectiveness of postharvest dip treatment with trifloxystrobin (TFX) or imazalil (IMZ) was compared for controlling green and blue mold (caused by Penicillium digitatum and Penicillium italicum, respectively) of citrus fruit. Residues retained by fruit were determined as a function of treatment time, dip temperature, and storage conditions. Trials on 'Avana apireno' mandarin oranges artificially inoculated with P. digitatum or P. italicum revealed that treatments with 200 to 600 mg/liter active ingredient TFX at 20 degrees C were less effective than 100 mg/liter TFX at 500C for controlling P. digitatum but equally effective for controlling P. italicum. IMZ treatments with 200 mg/liter IMZ at 20 degrees C or 25 mg/liter IMZ at 50 degrees C resulted in more than 98% reduction of P. digitatum and ca. 93% reduction of P. italicum compared with untreated fruit. Total suppression of pathogens was achieved when higher IMZ doses were applied. Studies on artificially wounded lemons, oranges, clementines, and mandarins revealed that treatment with 100 mg/liter TFX at 50 degrees C effectively controlled decay development (mainly due to P. digitatum) after 7 days of storage at 20 degrees C. These results were confirmed on nonwounded oranges of cv. Tarocco and on grapefruits of cvs. Marsh Seedless and Star Ruby during 3 weeks of simulated quarantine at 1 degrees C, storage (5 weeks at 8 degrees C for oranges and 8 weeks at 11degrees C for grapefruits), and an additional 1 week of simulated marketing conditions at 20 degrees C. IMZ at 50 degrees C was highly effective for controlling decay during storage and the simulated marketing period. TFX treatment at 50 degrees C was as effective as IMZ for controlling decay in most samples. After treatment with 100 mg/liter TFX at 20 degrees C, fungicide residues in 'Tarocco' oranges doubled from 0.15 mg/kg to 0.30 mg/kg when dip time increased from 0.5 to 3 min, whereas when treatments were performed at 50 degrees C TFX residues were not related to dipping time. Residues of TFX were significantly correlated with dip temperature. A 3-min dip treatment at 50 degrees C resulted in a deposition of TFX that was approximately twofold higher than that obtained when treatments were carried out at 20 degrees C.

Acetates↗

Aberrations of the tooth arcade and mandible in Dall's sheep from southwestern Yukon.

Nine Dall's sheep (Ovis dalli dalli) collected in the Ruby Mountains of southwestern Yukon were examined for aberrations of the tooth arcade and mandible. Missing and displaced teeth were common in older sheep (greater than or equal to 6 yr). Necrosis of the mandible was observed in five of eight rams and in one ewe. Corynebacterium pyogenes was cultured from exudates from two rams with enlarged fistulated mandibles. The sequence of events leading to lesions of the tooth arcade and to "lumpy jaw" is postulated to be associated with abnormal wearing of teeth and a disrupted alignment of the tooth arcade, impaction of vegetable matter between teeth and between the gingiva and the root of the tooth, and penetration of the traumatized gingiva membrane and alveolus by oral bacteria which induce a necrotizing infection.

Animal Feed↗

The politics of a renovation. Interview by Paul King and Donna Boss.

Recently, Shands Hospital at the University of Florida in Gainesville completed a $14-million foodservice renovation, one element in an 11-year-long, $100-million hospital-wide expansion project. Having successfully finished the work in her department, Food & Nutrition Services Director Ruby Puckett offers advice to her colleagues about how to stay in control of the renovation process from beginning through to the end.

Decision Making, Organizational↗

Total quality management: rhetoric and reality.

TQM in health care today is still more rhetoric than reality. Changing the typical health care organization's culture and installing the infrastructure needed to support TQM will be expensive and time consuming, and will require a new technology of health care quality control that is still developing. TQM will achieve its considerable potential only if management stays the course, invests the time and money--and reins in Ruby and her ilk.

Cardiology Service, Hospital↗

A cold nose can warm the heart.

Chase, a three-year-old golden retriever, is a show dog with a mission! For the past six months, Chase and his owner, Carol Ruby, of Orion Township, have been visiting patients on the inpatient Physical Rehabilitation Unit at Crittenton Hospital in Rochester. Chase played a key role in the creation of the unit's new Pet Therapy Program. Currently, Chase is one of three dogs whose goal is to bring smiles to the faces of patients at the hospital.

Animals↗

The piriform cortex and the endopiriform nucleus in the rat reveal generally similar pattern of connections.

The afferent and efferent connections of the piriform cortex and the endopiriform nucleus in the rat were studied by the method of axonal transport of two fluorescent tracers: Fluoro-Gold and Fluoro-Ruby. The results indicate that both structures possess not only the connections with the olfactory system, but also the reciprocal connections with the limbic system (entorhinal cortex, amygdaloid body), thalamus (mediodorsal and midline nuclei), extrapyramidal system (ventral part of the nucleus accumbens). The topographic differences in the organization of the association connections between the anterior and posterior parts of the piriform cortex are reported. Additionally, the reciprocal and relatively numerous connections between the endopiriform nucleus and the piriform cortex may result in their modulatory function, which in some pathological circumstances may have a critical significance in epileptogenesis.

Afferent Pathways↗

Corticostriatal projections from rat barrel cortex have an anisotropic organization that correlates with vibrissal whisking behavior.

To elucidate the detailed organization of corticostriatal projections from rodent somatosensory cortex, the anterograde tracers biotinylated dextran amine (BDA) and fluoro-ruby (FR) were injected into separate parts of the whisker "barrel" representation. In one group of rats, the two tracers were injected into different barrel columns residing in the same row; in the other group of rats, the tracers were deposited into barrel columns residing in different rows. Reconstructions of labeled axonal varicosities in the neostriatum and ventrobasal thalamus were analyzed quantitatively to compare the extent of overlapping projections to these subcortical structures. For both groups of animals, corticostriatal projections terminated in densely packed clusters that occupied curved lamellar-shaped regions along the dorsolateral edge of the neostriatum. When the tracers were injected into different whisker barrel rows, the distribution of BDA- and FR-labeled terminals in the neostriatum followed a crude somatotopic organization in which the amount of overlap was approximately the same as in the ventrobasal thalamus. When both tracers were injected into the same whisker barrel row, however, the amount of corticostriatal overlap was significantly higher than the amount of overlap observed in the ventrobasal thalamus. These results indicate that corticostriatal projections from whisker barrel cortex have an anisotropic organization that correlates with the pattern of vibrissal movements during whisking behavior.

Animals↗

Laser tattoo removal.

The availability of Q-switched ruby Nd:YAG and alexandrite lasers has revolutionized the treatment of tattoos. These modalities offer significant advantages over all previously available treatments and are currently the standard of care for the cosmetic removal of unwanted tattoos.

History, 20th Century↗

[Case report forum: the example of French sarcoma group multidisciplinary discussion tools. Sarcomes de la FNCLCC].

Within the Rubis 4th framework of European project is led a pilot experiment of tools and services for health professionals in prospect for the Aquitanian healthcare network. The sarcoma group of the FNCLCC (47 people) uses on its web site a multidisciplinary dialogue with a specific discussion forum. This service allows the anonymous publication of a imaging clinical case and to start a take care discussion. 87 cases were published in 13 months involving 261 answers from February 1999 to February 2000. A case is published every 4 days on average and the deadlines for replies regularly drop (15 days in February 1999 down 1.1 day in February 2000). The cases are published either for a diagnosis or treatment request (30%) or for the physical preparation of meeting or for the continuous medical training (70%). There are many advantages in comparison with the other possibilities of discussion: availability, autonomy of publication, cost, number of experts participating. These NTIC services will be developing within the regional healthcare oncology networks and are already tested by other regional groups (Lymphoma) considering the simplicity of use, management and training of the functionality.

Community Networks↗

Overlapping corticostriatal projections from the rodent vibrissal representations in primary and secondary somatosensory cortex.

To determine whether the neostriatum receives overlapping projections from two somatosensory cortical areas, the anterograde tracers biotinylated dextran amine (BDA) and fluoro-ruby (FR) were injected into the whisker representations of primary (SI) and secondary (SII) somatosensory cortex. Reconstructions of labeled terminals and their beaded varicosities in the neostriatum and thalamus were analyzed quantitatively to compare the extent of overlapping projections to both subcortical structures. Corticostriatal projections from focal sites in both somatosensory areas exhibited substantial amounts of divergence within the dorsolateral neostriatum. Most of the labeled terminals were concentrated in densely packed arborizations that occupied lamellar-shaped regions along the dorsolateral edge of the neostriatum. Tracer injections in both cortical areas also produced dense anterograde and retrograde labeling in the thalamus, especially in the ventrobasal complex (VB) and in the medial part of the posterior (POm) nucleus. Because these thalamic regions are topographically organized and have reciprocal connections with corresponding representations in both SI and SII, the amount of labeled overlap in the thalamus was used to indicate the degree of somatotopic correspondence at the SI and SII injection sites. We found that the proportion of overlapping projections to the neostriatum was moderately correlated with the amount of overlap observed in the thalamus. This result strongly indicates that specific sites in the dorsolateral neostriatum receive convergent projections from corresponding somatotopic representations in SI and SII, but also suggests that some of the corticostriatal divergence may reflect neostriatal integration of somatosensory information from noncorresponding representations in SI and SII.

Animals↗

Neural song control system of hummingbirds: comparison to swifts, vocal learning (Songbirds) and nonlearning (Suboscines) passerines, and vocal learning (Budgerigars) and nonlearning (Dove, owl, gull, quail, chicken) nonpasserines.

Males of certain hummingbird species such as Anna's hummingbirds (Calypte anna) learn their song during postnatal development. Here we report that male Anna's hummingbirds and male Amazilia hummingbirds (Amazilia amazilia), two singing hummingbird species, possess forebrain areas that are similar in morphological appearance, location, and connectivity to the song control areas RA (nucleus robustus archistriatalis), HVC (nucleus hyperstriatalis ventrale, pars caudale, or higher vocal center), and LMAN (lateral part of nucleus mangnocellularis anterioris) of oscine passerines (songbirds). The vocal control areas of songbirds are further defined by the expression of androgen receptors. Similarly, the singing hummingbird species express androgen receptors in the LMAN-like area and in the HVC-like area. The hummingbird RA projects to the medullary syringeal motonucleus nXIIts (nucleus hypoglossus pars tracheosyringealis) and the respiratory premotonucleus RAm (nucleus retroambigualis). The HVC-, RA-, and LMAN-like areas are rudimentary in adult male ruby-throated hummingbirds (Archilochus colubris) and Allen's hummingbirds (Selasphorus sasin) and not distinguishable in female hummingbirds, none of which sing. Vocal-area-like forebrain areas (delineated by the cytoarchitecture or androgen receptor expression) were not found in vocal nonlearning swifts and suboscines, the taxonomic sister groups of hummingbirds and songbirds, respectively. These areas were also missing in owls, ring doves, gulls, and gallinaceous species, nonpasserines that do not learn vocalizations. Budgerigars (vocal learners) are known to have forebrain vocal areas, but these areas do not express sex steroid receptors. These data suggest that hummingbirds and songbirds belong to two groups of birds that have common forebrain circuits. Parts of this circuit are organized as nuclear-like structures (LMAN, HVC, RA) in species that learn to sing.

Animals↗

Connectivity of the tectal zones coding for upward and downward oblique eye movements in goldfish.

Deep layers of the goldfish tectum code movements in a topographically ordered motor map. This work studies the relationship between tectal sites (coding eye movements with different vertical directions) and the distributions of boutons (left by their projections), within rostral mesencephalic structures and rhombencephalic reticular formations. These regions have been involved in the generation of the vertical and horizontal components of eye movement, respectively, as suggested by the Cartesian hypothesis of de-codification of tectal signal. With this aim, discrete injections of biotinylated dextran amine (BDA) and Fluoro-Ruby (FR) were made into functionally identified tectal sites, coding oblique eye movements with similar amplitude of the horizontal component but opposite upward and downward vertical directions, and the distribution of synaptic endings was determined. The main findings of the present work were as follows: 1) within the tectal descending tract, axons were organized according to the location of injected sites within the tectum; 2) BDA and FR boutons were distributed in separate clusters within the medial longitudinal fasciculus and oculomotor nuclei, as well as in the nearby mesencephalic reticular formation; and 3) the regions containing both types of bouton overlapped moderately within the mesencephalic reticular formation at the isthmus level. Overlapping was more extended at the different levels of the rhombencephalic reticular formation, although a shift in the distribution of both types of bouton was always observed. These results suggest that, within the vertical generator, the endings were separated to contact the different neuronal population that codes the upward and downward components of movements. In contrast, in the horizontal generator, tectal endings more likely converge on the same neuronal population to code the horizontal component of movements, irrespective of whether the oblique movements were directed upward or downward.

Animals↗

Overlapping corticostriatal projections from the rodent vibrissal representations in primary and secondary somatosensory cortex.

To determine whether the neostriatum receives overlapping projections from two somatosensory cortical areas, the anterograde tracers biotinylated dextran amine (BDA) and fluoro-ruby (FR) were injected into the whisker representations of primary (SI) and secondary (SII) somatosensory cortex. Reconstructions of labeled terminals and their beaded varicosities in the neostriatum and thalamus were analyzed quantitatively to compare the extent of overlapping projections to both subcortical structures. Corticostriatal projections from focal sites in both somatosensory areas exhibited substantial amounts of divergence within the dorsolateral neostriatum. Most of the labeled terminals were concentrated in densely packed arborizations that occupied lamellar-shaped regions along the dorsolateral edge of the neostriatum. Tracer injections in both cortical areas also produced dense anterograde and retrograde labeling in the thalamus, especially in the ventrobasal complex (VB) and in the medial part of the posterior (POm) nucleus. Because these thalamic regions are topographically organized and have reciprocal connections with corresponding representations in both SI and SII, the amount of labeled overlap in the thalamus was used to indicate the degree of somatotopic correspondence at the SI and SII injection sites. We found that the proportion of overlapping projections to the neostriatum was moderately correlated with the amount of overlap observed in the thalamus. This result strongly indicates that specific sites in the dorsolateral neostriatum receive convergent projections from corresponding somatotopic representations in SI and SII, but also suggests that some of the corticostriatal divergence may reflect neostriatal integration of somatosensory information from noncorresponding representations in SI and SII.

Animals↗