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Aberrant gene expression at the creatine kinase loci during Barbus hybrid development (Cypriniformes, Teleostei).

The tissue specificity and ontogeny of creatine kinase (CK, EC 2.7.3.2) are reported for tiger, rosy, ruby, cherry and gold barbs, and in interspecific hybrids where the tiger barb is the maternal parent. The spatial expression of CK isoenzymes in Barbus is consistent with the tissue patterns reported in other teleosts. In general, as the taxonomic (genetic) distance between the parental species increases, a corresponding delay in embryonic gene expression occurs; suggestive of species-specific effector molecule/sensor gene induction thresholds.

Animals↗

The basal forebrain projection to the region of the nuclei gemini in the rat; a combined light and electron microscopic study employing horseradish peroxidase, fluorescent tracers and Phaseolus vulgaris-leucoagglutinin.

We have examined the location of basal forebrain cells projecting to the region of the nuclei gemini in the caudolateral hypothalamus of the rat using retrograde transport of wheatgerm agglutinin-horseradish peroxidase. Since many tracer-positive neurons were identified in ventral pallidal areas known to project to the mediodorsal nucleus of the thalamus, we also prepared several animals with wheatgerm agglutinin-horseradish peroxidase injections in mediodorsal thalamus. Many of the sections from both groups of animals were subsequently prepared for the demonstration of ventral pallidal regions, using either substance P or glutamate decarboxylase as a pallidal marker. Some animals received injections of different retrogradely transported fluorescent tracers in the mediodorsal thalamus and the nuclei gemini for the purpose of studying potential axon collateralization. The large gemini-projecting cells are diffusely scattered within the medial forebrain bundle area, from the caudal margin of the nucleus of the horizontal limb of the diagonal band to the rostral tip of the olfactory tubercle, and with a concentration of cells in the lateral part of the medial forebrain bundle region. Gemini-projecting cells were not found in the olfactory tubercle proper, including the islands of Calleja complexes, or in the ventral pallidal areas located dorsal to the medial forebrain bundle area underneath the lateral extension of the anterior commissure. Gemini-projecting cells within ventral pallidal areas were observed only in regions where the longitudinal fascicles of the medial forebrain bundle interdigitate with the rostroventral extension of the ventral pallidum. Anterogradely-labeled fiber plexuses in the region of the nuclei gemini were observed following injection of Phaseolus vulgaris-leucoagglutinin or Fluoro-Ruby into the forebrain regions containing retrogradely-labeled neurons following nuclei gemini injections of wheatgerm agglutinin-horseradish peroxidase. We found no evidence of cells with axonal projections to both mediodorsal thalamus and nuclei gemini. The gemini-projecting cells are generally large, triangular and plump, and the electron microscopic picture of gemini-projecting neurons is the same regardless of whether the cells are located in pallidal or non-pallidal areas.

Animals↗

Anatomical plasticity in the medial superior olive following ablation of the inferior colliculus in neonatal and adult rats.

We evaluated the consequences of unilateral ablation of the inferior colliculus (IC) upon the ascending projection from the medial superior olive (MSO) to the IC. Ablation of the IC was performed in rats aged between postnatal day 1 (P1) and maturity. All the rats were given injections of Fluoro-Gold (FG) into the ipsilateral IC at birth (P0) (before the ipsilateral IC was ablated in any case) so that growth of early-developing axons to the ipsilateral IC could be examined for any labeled neurons in the ipsilateral MSO. Upon reaching adulthood, the rats received injections of Fluoro-Ruby (FR) into the contralateral (intact) IC so that aberrant crossed projections to the intact IC could be examined for any labeled neurons in the ipsilateral MSO. These rats were killed 2 days after FR injections. The number of surviving cells in the ipsilateral MSO were counted in Nissl-stained sections for quantitative analysis of retrograde degeneration. The results show that: (1) the total number of neurons was reduced to 64-34% in the ipsilateral MSO as a result of IC ablation; (2) cell reduction by retrograde degeneration followed a U-shaped curve with a maximal effect in rats operated at P7 (reduced to 34%); (3) adult ablation of the IC led to retrograde degeneration that was less severe than that in late neonatal (P7) ablation; (4) an aberrant projection from the MSO to the contralateral IC occurred in rats operated at P1 and P3 but not in rats operated at P7 or maturity. Thus, our findings suggest that growth of late-developing axons is a major factor in the plasticity of this system of projection.

Age Factors↗

Quantitative assessment of the ability of collateral sprouting of the motor and primary sensory neurons after the end-to-side neurorrhaphy of the rat musculocutaneous nerve with the ulnar nerve.

In view of the Lack of theoretical information, end-to-side neurorrhaphy is a frequent object of experimental interest. End-to-side neurorrhaphy is based on collateral sprouting of an intact axon. The quantitative assessment of collateral sprouts sent by an intact motor and sensory axon was the goal of the present study. End-to-side neurorrhaphy of the distal stump of transected musculocutaneous nerve (MCN) with intact ulnar nerve (UN) was performed in a rat model. Collateral sprouts were quantitatively evaluated by counting of motoneurons and DRG neurons following their retrograde labeling by Fluoro-Ruby and Fluoro-Emerald applied to the UN and MCN, respectively. The results suggest a comparable capacity of both intact sensory and motor axons to send collateral sprouts into a denervated nerve stump. The ratio of sensory/motor neurons, the axons of which reinnervated distal MCN stumps, was very similar to that of intact UN (6.500 and 6.747, respectively), but different from intact MCN (5.029). This suggests that the pruning process occurred to balance the collateral sprouts at a ratio of sensory/motor neurons for the donor UN, but not according to the number of sensory and motor bands of Bungner available in the distal stump of the MCN. The present experimental study confirms end-to-side neurorrhaphy as a suitable method of nerve reconstruction.

Anastomosis, Surgical↗

Maternal vasculopathy and histologic diagnosis of preeclampsia: poor correlation of histologic changes and clinical manifestation.

OBJECTIVE: The purpose of this study was to evaluate the frequency of maternal vasculopathy and to determine the usefulness of placental examination in correlation with clinical manifestation of pregnancy-induced hypertension. STUDY DESIGN: We investigated 350 pregnancy-induced hypertension placentas over a 2.5-year period (January 2002-June 2004) to identify the potential usefulness of the maternal vasculopathy in histologic diagnosis and corroboration of clinical preeclampsia. This study was carried out at West Virginia University Ruby Memorial Hospital, where 1271 placentas were examined macroscopically and microscopically for the period. The presence or absence of vascular changes, infarctions, thrombosis, abruption, chorangiosis, and chorioamnionitis were investigated for each placenta with the clinical diagnosis of preeclampsia or pregnancy-induced hypertension. RESULTS: Two types of vascular changes that are characteristic of pregnancy-induced hypertension, namely maternal atherosis and fibrinoid medial necrosis, were found in 21.4% of the placentas (75/350 placentas). These vascular changes were found mainly in the decidua capsularis of the membrane rolls, not on the decidua basalis of the maternal floor. There were more placental infarctions within these placentas (32.3%; 113/350 placentas). Intervillous thrombosis was 18.9% (66/350 placentas); microscopic abruption was 6.3% (22/350 placentas); chorangiosis was 10.9% (38/350 placentas), and chorioamnionitis was 8.6% (30/350 placentas). CONCLUSION: These findings demonstrate that the maternal vascular changes that are characteristic of pregnancy-induced hypertension are found in approximately 1 of 5 pregnancy-induced hypertension placentas. Examination of pregnancy-induced hypertension placentas should be based on a combination of a number of histologic changes. Furthermore, the decidua capsularis of the fetal membrane roll, rather than the decidua basalis of the maternal floor, of the placenta should be examined for these maternal vascular changes.

Female↗

Theoretical estimates of the solid angle subtended by a dual diaphragm-detector assembly for alpha sources.

Knowledge of the solid angle subtended by a detector is essential for the determination of the activity of a radioactive source. This work investigates the influence of diaphragm and source diameter, and source to diaphragm distance, on the solid angle subtended by a dual opposed alpha diaphragm-detector assembly. Expressions published by Segre, Ruby, and others, and the Monte Carlo method are used. The analytical expressions consistently produced higher estimates than the Monte Carlo method for the solid angle.

Journal Article↗

A comparison of different analytical methods of determining the solid angle of a circular coaxial source-detector system.

In this work, seven different analytical expressions for the solid angle subtended by a circular detector for a coaxial circular source are compared. These expressions -- which have been published by different investigators over the course of the last half century -- are compared for previously published geometries, and for geometries planned in future experimental work. As compared to benchmark Monte Carlo calculations, the expressions published by Segre, Knoll, and Ruby, were the most accurate and exhibited the highest degree of self-correlation.

Journal Article↗

The solid angle subtended by a circular detector for a linear source.

An exact solution is presented for the solid angle subtended by a circular detector for a linear source. It is based on Conway's equation for a non-coaxial point source, involving an integral of Bessel functions [Conway, J.T., 2006. Generalizations of Ruby's formula for the geometric efficiency of a parallel-disk source and detector system. Nucl. Instrum. Methods A 562, 146-153]. Some numerical examples are calculated and compared with the results obtained with a recently published closed formula (Galiano, E., Pagnutti, C., 2006. An analytical solution for the solid angle subtended by a circular detector for a symmetrically positioned linear source. Appl. Radiat. Isot. 64, 603-607), which is incorrect but can be improved by applying a correction factor.

Journal Article↗

Plasticity of vagal afferents at the site of an incision in the wall of the stomach.

Our objectives were to determine whether the vagal afferent innervation of the stomach reorganizes after surgery and to observe how different wound closure techniques might influence such a process. The smooth muscle wall of the stomach served as a model because it is densely innervated by vagal axons and is frequently compromised by gastric surgery. Male Sprague-Dawley rats were assigned to one of six groups: three groups served as controls in which the stomach was exposed surgically and a) subjected to no further manipulation, b) traumatized with suture needle punctures of the muscle wall, or c) insulted by the placement of knotted suture thread in the stomach muscle; three surgical groups received a 1.0 cm incision through the ventral muscle wall of the stomach that was closed using either a) absorbable sutures, b) fibrin glue, or c) n-butyl cyanoacrylate. Rats were killed 4 to 7 months post-surgery. Prior to euthanasia, Micro-Ruby was injected into the left nodose ganglion of each rat to label vagal afferent axons and terminals. Twelve days post-injection, the stomachs were processed for microscopy. All groups recovered quickly from surgery, without differences in body weight. The presence of suture material in the muscle wall of the stomach was sufficient to produce reorganization of nearby vagal afferents. In addition, we observed that an incision of the smooth muscle wall of the stomach and the associated damage to vagal afferents provoked reorganization and regeneration of vagal afferents. Vagal remodeling at the incision was characteristic of axonal patterns found in neuromas (unlike the organized regeneration and differentiation that can occur after axotomy central to the target organ). Vagal afferent terminals located at the site of the incision were free nerve endings and growth cone profiles, and not the characteristically complex end organs normally found in the smooth muscle. Finally, the pattern of vagal plasticity was influenced by the wound closure technique used. Overall, the remodeling of afferents was aberrant in nature, and such neural pathology could contribute to the neuropathic symptoms and hyperalgesias associated with gastrointestinal trauma and bariatric surgery.

Animals↗

Phosphoproteomic profiling of human SH-SY5Y neuroblastoma cells during response to 6-hydroxydopamine-induced oxidative stress.

Dopaminergic neurons are known to be vulnerable to age-related neuronal disorders due to reactive oxygen species (ROS) generated during dopamine metabolism. However, it remains unclear what kinds of proteins are involved in the response to oxidative stress. We examined changes in whole proteins and phosphoproteins in the human dopaminergic neuroblastoma cell line SH-SY5Y under oxidative stress induced by the dopaminergic neurotoxin 6-hydroxydopamine (6-OHDA). Proteins of SH-SY5Y cells at various stages of oxidative stress were separated by two-dimensional gel electrophoresis for comparative analysis. Increase in glutathione-S-transferase pi was detected on SYPRO Ruby-stained gels by computer-aided image analysis. Stress-induced alterations in phosphoproteins were detected by Pro-Q Diamond staining. Elongation factor 2, lamin A/C, T-complex protein 1, and heterogeneous nuclear ribonucleoprotein H3 were identified by MALDI-TOF mass spectrometry as stress-responsive elements.

Blotting, Western↗

Development of a low-pressure diamond anvil cell and analytical tools to monitor microbial activities in situ under controlled P and T.

We have designed a new low-pressure Diamond Anvil Cell (DAC), calibrated two novel pressure calibrants and validated the use of semi-quantitative Raman and X-ray spectroscopies to monitor the fate of microbes, their metabolism or their cellular components under controlled pressures and temperatures in the 0.1-1.4 GPa and 20-300 degrees C P,T range. The low-pressure DAC has a 250- to 600-microm-thick observation diamond window to allow for lower detection limits and improved microscopic imaging. This new design allows the determination of cellular growth parameters from automated image analysis, which can be correlated with the spectroscopic data obtained on metabolism, ensuring high quality data collection on microbial activity under pressure. The novel pressure sensors offer the ease of use of the well-known ruby scale, while being more sensitive and reacting to pressure variations instantaneously.

Bacteria↗

Hydrogen peroxide increases a 55-kDa tyrosinase concomitantly with induction of p53-dependent p21 waf1 expression and a greater Bax/Bcl-2 ratio in pigmented melanoma.

Differentiated melanocytic cells produce melanin, through several redox reactions including tyrosinase-catalyzed DOPA oxidation to DOPA quinone. We now developed a method based on DOPA oxidase in-gel detection and Sypro Ruby fluorometric normalization to investigate induction of specific DOPA oxidase isoforms in response to hydrogen peroxide-mediated stress, and to ask whether this is associated with p53-dependent adaptive responses. This report shows that hydrogen peroxide leads to comparable induction of 60 and 55 kDa DOPA oxidases in poorly pigmented B16 melanoma, in contrast to sole induction of a major 55 kDa DOPA oxidase in their highly pigmented counterparts. In the latter cells, this response also increases p53 concomitant with joint induction of p53-activated proteins like the cell-cycle inhibitor p21WAF1 and pro-apoptotic bax, with no comparable effect on expression of anti-apoptotic bcl-2. Together, these data suggest that response to hydrogen peroxide involves p53-mediated growth-restrictive signaling and unequal induction of specific DOPA oxidases in melanocytic cells with unequal basal pigmentation.

Animals↗

The effect of support material and conditioning on wettability of PAH/PSS multilayer films.

An efficient method for characterizing wetting properties of heterogeneous surfaces produced by sequential adsorption of polyelectrolytes was developed. Three types of polyelectrolytes were used: polyallylamine hydrochloride (PAH), polyethyleneimine (PEI), both of a cationic type, and polysodium 4-styrenesulfonate (PSS), of an anionic type. Multilayer films were prepared by 'layer-by-layer' (LbL) deposition technique. Natural ruby mica, glass, titanium foil and silicon wafers were used as the support material for PE films. Wetting of polyelectrolyte films was determined experimentally by contact angle measurements, using technique of direct image analysis of shape of sessile drops. Periodic oscillations in contact angle values were observed for multilayers terminated by polycation and polyanion, respectively, and the variations in contact angle values strongly depended on the conditions of adsorption and multilayer treatment after deposition. Therefore, the influence of ionic strength of polyelectrolyte solution used for deposition on wetting of multilayer films was considered and also the effect of conditioning in different environments was investigated. It is usually assumed that film properties and stability strongly depend on the first layer which is used to anchor a multilayer at the surface of support material. To investigate influence of the first layer, PAH/PSS films were compared with more complex ones having PEI as the first layer with a sequence of PSS/PAH deposited on top of it.

Adsorption↗

Study of proteomic changes associated with healthy and tumoral murine samples in neuroblastoma by principal component analysis and classification methods.

BACKGROUND: The adrenal gland is the election organ forming primary neuroblastoma (NB) tumours, the most common extracranial solid tumours of infancy and childhood. METHODS: Samples of adrenal gland belonging to healthy and diseased nude mouse were analysed by 2D gel-electrophoresis. The resulting 2D-PAGE maps were digitized by PDQuest and investigated by principal component analysis (PCA). RESULTS: The analysis of the loadings of the first principal component (PC) permitted the evaluation of the spots characterising each class of samples. Moreover, the soft-independent model of class analogy (SIMCA) method confirmed the separation of the samples in the two classes and allowed the identification of the modelling and discriminating spots. Very good correlation was found between the data obtained by analysis of 2D maps via the commercial software PDQuest and the present PCA analysis. In both cases, the comparison between such maps showed up- and down-regulation of 84 polypeptide chains, out of a total of 700 spots detected by a fluorescent stain, Sypro Ruby. Spots that were differentially expressed between the two groups were analysed by matrix-assisted laser desorption time-of-flight (MALDI-TOF) mass spectrometry and 14 of these spots were identified so far.

Animals↗

Influence of ozonation on extractability of Pb and Zn from contaminated soils.

The effect of soil ozonation on Pb and Zn extraction with EDTA, bioavailability (Ruby's Physiologically Based Extraction Test, PBET) and mobility (Toxicity Characteristic Leaching Procedure, TCLP) of Pb was studied on contaminated soils taken from 7 different locations in the Mezica Valley, Slovenia. EDTA extraction (40 mmol kg(-1)) removed from 27.4+/-1.5% to 64.8+/-1.5% of Pb, and from 1.9+/-0.2% to 22.4+/-2.0% of Zn from tested soils, and significantly reduced soil Pb bioavailability (PBET) and mobility (TCLP). Pretreatment of tested soils with ozone before EDTA extraction enhanced EDTA extractability of Pb for 11.0 to 28.9%, but had no effect on the extractability of Zn. In most of the soils, ozonation had no statistically significant effect on bioavailability and mobility of Pb, residual after EDTA extraction. Using linear regression analysis we found a significant increase (p<0.01) in EDTA extractability of Pb after soil ozonation in soils with a higher initial Pb content. EDTA extractability of Pb after soil ozonation was also significantly higher for soils with a lower Pb extractability when treated with EDTA alone. We found no correlation between soil organic matter content and the percentage of the Pb fraction bound to soil organic matter (where from 25.6+/-1.3% to 73.2+/-0.6% of Pb reside in tested soils) and Pb extractability with EDTA after soil ozonation.

Biological Availability↗

Laser treatment of tattoos.

Tattooing has been around since the early beginnings of modern civilization. Modern tattoo artists use a myriad of colors to produce striking designs, resulting in permanent works of body art; however, we humans have been changing our minds since the beginning of time. Our fickle nature results in the desire to change what has been placed as a permanent reminder of a friend, spouse, or work of art. Removing tattoos began with abrasive and destructive measures to destroy the tattoo, and unfortunately, the skin it was contained in. The discovery of selective photothermolysis, the ability to selectively remove target structures without disrupting the surrounding skin, made it at least possible to remove tattoos without destroying the surrounding skin leaving a scar. Theory predicted that pulse durations in the nanosecond domain would be optimal for tattoo removal, and the Q-switched neodymium:yttrium-aluminum-garnet, alexandrite, and ruby lasers fulfilled this need. Too often, older lasers or intense pulsed light sources are used to treat tattoos, often with significant scarring. Since the advent of the Q-switched lasers more than a decade ago, improvement in tattoo-removal lasers has been incremental. Developments leading to new tattoo inks, feedback systems to detect the absorbance characteristics of tattoo inks, dermal clearing agents, and perhaps even shorter pulse-duration lasers should result in improved results for the future.

Humans↗

Identification of major cellular proteins synthesized in response to interleukin-1 and interleukin-6 in human hepatoma HepG2 cells.

Interleukin-1 (IL-1) and interleukin-6 (IL-6) are principal proinflammatory cytokines inducing the acute phase response of various tissues, including liver. Cultured human hepatoma HepG2 cells were stimulated with IL-1 (10 ng/ml) and IL-6 (10 ng/ml). After 24 h the cells were collected and disrupted by sonication in a lysis buffer containing 8M urea. The extracted cellular proteins were separated by 2D polyacrylamide gel electrophoresis. The gels were stained with Coomassie Brilliant Blue R-250 and the protein spots showing different intensities in comparison to control (unstimulated) cells were excised and subjected to analysis by LC-MS/MS. Alternatively, proteins were stained with SYPRO Ruby. These differentially expressed proteins include seven up-regulated and two down-regulated intracellular proteins of various functions. The identification of three cytokine-responsive proteins was confirmed by biosynthetic labeling with [35S]methionine after incubation of HepG2 cells, and by western blot with specific antisera.

Actin Depolymerizing Factors↗

Effect of the applied load on surface contact fatigue of dental filling materials.

OBJECTIVE: The purpose of this study was to investigate the effect of the applied load in a rolling-ball device on the surface contact fatigue of two dental filling materials: a microfilled composite (FA) and a glass ionomer (GFII). METHODS: Disk specimens 10 mm diameter by 1.5 mm thick were set into cavities cut in plates of PMMA. After setting, the specimens were ground and polished using wet-carborundum paper followed by 1 microm alumina and then stored for 24 h in water at 23 degrees C. The surface fatigue test was carried out using loads ranging from 100 to 500 gf through a ruby ball 2 mm diameter using a rolling-ball device. The ball was set to rotate at 720 rpm and a surface profile was determined on impressions recorded at regular intervals. Fatigue life was defined as the number of cycles to produce a track 5 microm deep and was determined by interpolation. Data were statistically analyzed by ANOVA and t-tests at a significance level of 0.05. RESULTS: At loads ranging from 200 to 500 gf the fatigue life was 0.7-10.6 x 10(5) cycles for FA, and 0.7-4.8 x 10(3) cycles for GFII. For both materials the fatigue life was reduced significantly by increasing the test load (p<0.05). The surface contact fatigue life for FA was at least 100 times greater than that for GFII at all loads. SIGNIFICANCE: The rolling ball device was confirmed as a simple and meaningful device for the study of surface contact fatigue. A microfilled composite is very much more resistant to contact fatigue than a glass ionomer.

Composite Resins↗