Reduction of erosion by protein-containing toothpastes.

AIM: To assess the effect of protein-containing toothpastes on the progression of dental erosion in situ (with pellicle) and in vitro (without pellicle). METHODS: A combined split-mouth (extraoral water or toothpaste brushing) and crossover (type of toothpaste) setup was used. Two protein-containing (high/low concentrations of colostrum) and one nonprotein (placebo) toothpaste were investigated. Sixteen volunteers wore intraoral appliances containing 2 human enamel samples on 3 afternoons for pellicle growth during 90 min. One enamel sample was brushed for 5 s with one of the three toothpastes and subsequently exposed to a slurry of the corresponding toothpaste for 2 min. The other sample was exposed to water. Both samples were subsequently exposed to citric acid (extraorally). Loss of calcium and inorganic phosphate were determined. The same sequence of exposures was applied to 16 enamel samples in an in vitro setup without pellicle. RESULTS: With the in situ-formed pellicle, all toothpastes significantly reduced calcium loss compared to water brushing, although no significant differences were found among toothpastes (p = 0.073). For the loss of phosphate, a significant reduction could be found with the use of the high-protein toothpaste compared to the nonprotein toothpaste. Overall there were only slight differences between the toothpastes. Toothpaste effects were less clear in the in vitro experiment. CONCLUSION: The addition of proteins to toothpaste shows some promise for the prevention of erosion. Further research is needed to investigate the performance of the protein-containing toothpastes in longer in situ studies with regard to erosive wear.

Fluid-structure effects of cloaking a submerged spherical shell.

Backscattering from a cloaked submerged spherical shell is analyzed in the low, mid, and high frequency regimes. Complex poles of the scattered pressure amplitudes using Cauchy residue theory are evaluated in an effort to explain dominant features of the scattered pressure and how they are affected by the introduction of a cloak. The methodology used is similar to that performed by Sammelmann and Hackman [J. Acoust. Soc. Am. 85, 114-124 (1989); J. Acoust. Soc. Am. 89, 2096-2103 (1991); J. Acoust. Soc. Am. 90, 2705-2717 (1991)] in a series of papers written on scattering from an uncloaked spherical shell. In general, it is found that cloaking has the effect of diminishing the amplitude and shifting tonal backscatter responses. Extreme changes of normal and tangential fluid phase velocities at the fluid-solid interface when cloaking is employed leads to elimination of the "mid-frequency enhancement" near the coincidence frequency for even modestly effective cloaks, while reduction of the "high-frequency enhancement" resulting from the "thickness quasi-resonance" near the cut-off frequency of the symmetric (S2(B)) mode requires more effective cloaking, but can be practically eliminated by employing a cloak that creates tangential acoustic velocities in excess of the S2(B) mode phase speed near cutoff.

A new polymorphism in the Growth and Differentiation Factor 9 (GDF9) gene is associated with increased ovulation rate and prolificacy in homozygous sheep.

Brazilian Santa Inês (SI) sheep are very well-adapted to the tropical conditions of Brazil and are an important source of animal protein. A high rate of twin births was reported in some SI flocks. Growth and Differentiation Factor 9 (GDF9) and Bone Morphogenetic Protein 15 (BMP15) are the first two genes expressed by the oocyte to be associated with an increased ovulation rate in sheep. All GDF9 and BMP15 variants characterized, until now, present the same phenotype: the heterozygote ewes have an increased ovulation rate and the mutated homozygotes are sterile. In this study, we have found a new allele of GDF9, named FecG(E) (Embrapa), which leads to a substitution of a phenylalanine with a cysteine in a conservative position of the mature peptide. Homozygote ewes presenting the FecG(E) allele have shown an increase in their ovulation rate (82%) and prolificacy (58%). This new phenotype can be very useful in better understanding the genetic control of follicular development; the mechanisms involved in the control of ovulation rate in mammals; and for the improvement of sheep production.

In vitro reduction of dental erosion by low-concentration TiF4 solutions.

The aims of this study were to compare daily versus single applications of low-concentration TiF(4) solutions for reduction of enamel erosion and to evaluate the enamel surface loss due to application of these solutions. Sixty bovine enamel samples were randomly divided into 2 groups: single versus daily treatment with TiF(4) solution (ST vs. DT), which were subdivided into 5 subgroups (n = 6): 0% (control); 0.1, 0.5, 0.75 and 1% TiF(4) concentration. Fluoride treatment was performed by immersing specimens in 10 ml of the TiF(4) solutions for 5 min and rinsing them with tap water for 30 s. ST specimens were treated once only, DT specimens were treated before each erosion cycle. All specimens were subjected to 4 erosion cycles: 6 immersions of 2 min in Sprite, tap water rinse for 1 min and storage in artificial saliva for 1.5 h. Erosive enamel loss was measured using light profilometry, after each fluoride treatment and each erosive cycle. The values of surface loss/gain at application of the TiF(4) solutions did not significantly differ from zero. At the end of the 4 erosion cycles, 0.5% showed the least (ST: 3.45 ± 0.27 µm, DT: 1.08 ± 1.69 µm) and 1% showed the most surface loss (ST: 4.87 ± 1.13 µm, DT: 6.56 ± 1.49 µm). A significant reduction of surface loss was found only for 0.5% DT (p = 0.009). Within the limitations of an in vitro study, it was concluded that multiple applications of a 0.5% TiF(4) solution significantly reduced enamel erosion in vitro and caused no enamel loss at application.

Effect of salivary factors on the susceptibility of hydroxyapatite to early erosion.

OBJECTIVE: Salivary pellicle is known to reduce the erosion of enamel and differences in the level of protection exist between individual saliva sources, but which parameters or components are important is not known. The focus of this study was to investigate the relationship between saliva parameters and early erosion of hydroxyapatite (HAp) with an in situ grown saliva film. METHODS: Twenty-eight volunteers carried two HAp and one porcelain discs in their buccal sulcus for 1.5 h. Next, the discs covered with pellicle and the attached saliva film were exposed extraorally to 50 mM (pH = 3) citric acid for 2 min and unstimulated and stimulated saliva was collected. Calcium loss from HAp after erosive challenge was measured, corrected for calcium loss from pellicle on porcelain discs and averaged. Several salivary parameters were analysed. Pearson's linear correlation and multiple regression analysis were used to study the relation between saliva parameters and HAp erosion. RESULTS: Significant correlations were found between HAp erosion and the concentration of phosphorus in unstimulated saliva (r = 0.40, p = 0.03) and between HAp erosion and the concentration of sodium (r = -0.40, p = 0.03), chloride (r = -0.47, p = 0.01), phosphorus (r = 0.45, p = 0.01) and flow (r = -0.39, p = 0.04) of stimulated saliva. Multivariate analysis revealed a significant role in the HAp erosion for sodium, urea, total protein, albumin, pH and flow of unstimulated saliva and for sodium, potassium, urea, and phosphorus of stimulated saliva. CONCLUSIONS: Several salivary parameters are associated with the susceptibility of HAp to erosion.

Reduction of erosive wear in situ by stannous fluoride-containing toothpaste.

BACKGROUND/AIMS: Stannous fluoride (SnF) has been suggested as a dental erosion-preventive agent. The aim of this single-centre, randomized, double-blind, in situ study was to evaluate the effect of toothpastes with SnF in the prevention of erosive enamel wear. METHODS: A combined split-mouth (extra-oral water or toothpaste brushing) and crossover (type of toothpaste) set-up was used. Twelve volunteers wore palatal appliances containing human enamel samples. Three toothpastes were used, in three consecutive runs, in randomized order: two toothpastes containing SnF (coded M and PE) and one toothpaste containing only sodium fluoride (coded C). On day 1 of each run the appliances were worn for pellicle formation. On days 2-5 the samples were also brushed twice with a toothpaste-water slurry or only water (control). Erosion took place on days 2-5 extra-orally 3 times a day (5 min) in a citric acid solution (pH 2.3). Enamel wear depth was quantified by optical profilometry. The effect of toothpastes was tested using General Linear Modeling. RESULTS: Average erosive wear depth of control samples was 23 µm. Both SnF toothpastes significantly reduced erosive wear: M by 34% (SD 39%) and PE by 26% (SD 25%). The control toothpaste reduced erosive wear non-significantly by 7% (SD 20%). Both SnF-containing toothpastes significantly reduced erosive wear compared to the sodium fluoride toothpaste. CONCLUSION: We conclude that SnF-containing toothpastes are able to reduce erosive tooth wear in situ.

Myocardial ischemia during endoscopic retrograde cholangiopancreatography - A real problem?

AIM: The occurrence of silent myocardial ischemia during endoscopic retrograde cholangiopancreatography (ERCP) has been documented, but its clinical significance remains unknown. The aims of this study were to investigate the incidence and risk factors of myocardial ischemia during ERCP, to determine the presence or absence of permanent myocardial injury and to evaluate if deep sedation with propofol had a positive effect on myocardial ischemia during ERCP. METHODS: Ambulatory ST-segment monitoring from 30 minutes prior to 4 hours after ERCP was obtained on 50 patients. A deep sedation was performed with intravenous propofol administered by anesthesiologist. Changes in vital signs during ERCP, pre and postprocedural 12-lead ECG examination and cardiac enzymes were evaluated. RESULTS: Silent cardiac ischemia occurred only in one patient (2%) during ERCP. This 64-year-old patient did not develop hypoxemia, tachycardia or hypotension periods during the exam. None of the patients developed cardiac enzymes or postprocedural electrocardiographic changes. Thirty seven (74%) patients suffered rhythm changes. CONCLUSION: Although rhythm disturbances were common, silent myocardial ischemia during ERCP was rare (2%) and without clinical relevance. In prolonged or complex therapeutic procedures, like ERCP, deep sedation with propofol performed by trained personnel is associated with reduced cardiac complications.

Preslaughter mortality of broilers in relation to lairage and season in a subtropical climate.

The preslaughter handling and transport of broilers are stressful operations that might affect welfare and meat quality and could increase numbers of deaths before slaughter. However, the influence of thermal factors during transportation and lairage at slaughterhouses is complex in subtropical regions, where increasing temperature and high RH are the major concerns regarding animal survival before slaughter. In this study we assessed the influence of a controlled lairage environment on preslaughter mortality rates of broiler chickens that were transported during different seasons of the year and had varying lairage times in the subtropical climate. Preslaughter data from 13,937 broiler flocks were recorded daily during 2006 in a commercial slaughterhouse in southeastern Brazil. The main factors that influenced daily mortality rate were mean dry bulb temperature and RH, lairage time, daily periods, density of broilers per crate, season of the year, stocking density per lorry, transport time, and distance between farms and slaughterhouse. A holding area at the slaughterhouse with environmental control was assessed. Using a double GLM for mean and dispersion modeling, the seasons were found to have significant effects (P < 0.05) on average mortality rates. The highest incidence was observed in summer (0.42%), followed by spring (0.39%), winter (0.28%), and autumn (0.23%). A decrease of preslaughter mortality of broilers during summer (P < 0.05) was observed when the lairage time was increased, mainly after 1 h of exposure to a controlled environment. Thus, lairage for 3 to 4 h in a controlled lairage environment during the summer and spring is necessary to reduce the thermal load of broiler chickens.

The importance of ultrasound findings in the study of anal pain.

OBJECTIVE: Endoanal ultrasonography can detect organic causes of anal pain without pathology on physical examination. The aim of this study is to evaluate the importance of endoanal ultrasonography in the diagnosis and therapeutic management of idiopathic and functional anal pain. MATERIAL AND METHODS: Retrospective study, between 15 March 2005 and 15 June 2008, of all patients with proctalgia and normal examination or with alterations not responsible for anal pain at proctologic exam that have undergone an endoanal ultrasonography. RESULTS: A total of 90 patients were analyzed, with a mean age of 50.5 years, 58% were female. Twenty-three patients had functional anal pain clinic criteria. Endoanal ultrasonography revealed alterations in 49% of patients. The primary findings were changes in sphincters in 14 patients, followed by anal sepsis in 12 patients, anal fissure in 10 patients, perirectal lesions in 6 patients and ulcer of the anal canal in 2 patients. Of the patients with sphincter defects, 5 patients had criteria of chronic anal pain. In this group of patients, no differences were found in manometric and defecographic results between the different ultrasound abnormalities. CONCLUSIONS: The endoanal ultrasonography detected occult organic lesions to proctologic examination, in half the patients with anal pain. Ultrasound abnormalities were found in 22% of patients with functional anal pain. However, there was no correlation between ultrasound findings and physiological studies, and therefore could not find etiological or pathogenic factors of functional anal pain.

Predicting Rice Stem Stink Bug Population Dynamics Based on GAMLSS Models.

The rice stem stink bug, Tibraca limbativentris Stål (Hemiptera: Pentatomidae), is one of the most harmful insects for Brazilian rice fields. Aiming to define the most appropriate time and place for pest management measures in commercial paddy fields, we adjusted regression models (Poisson, Zero Inflated Poisson, reparametrized Zero Inflated Poisson, Negative Binomial and Zero Inflated Negative Binomial) for modeling the population variation of T. limbativentris along the phenological cycle of the flooded rice cultivation. We hypothesize that the rice stem stink bug population's size is influenced by the rice cycle (time) and geographical positions within the crop. It was possible to predict the occurrence of the rice stem stink bug in the commercial flooded rice crop. The population of the rice stem stink bug increased significantly with the time or phenological evolution of rice. Our results indicated that the start of T. limbativentris monitoring should occur up to 45 d After Plant Emergence (DAE), from the regions along the edges of the rice paddies, which are the points of entry and higher concentration of the insect. In addition, 45 and 60 DAE were considered the crucial times for T. limbativentris control decision making in flooded rice paddies.

ATL-1, an analogue of aspirin-triggered lipoxin A4, is a potent inhibitor of several steps in angiogenesis induced by vascular endothelial growth factor.

BACKGROUND AND PURPOSE: Vascular endothelial growth factor (VEGF) is the most important proangiogenic protein. We have demonstrated that ATL-1, a synthetic analogue of aspirin-triggered lipoxin A(4), inhibits VEGF-induced endothelial cell (EC) migration. In the present study, we investigated the effects of ATL-1 in several other actions stimulated by VEGF. METHODS: Human umbilical vein ECs were treated with ATL-1 for 30 min before stimulation with VEGF. Cell proliferation was measured by thymidine incorporation. Adherent cells were determined by fluorescence intensity using a Multilabel counter. Expression and activity of matrix metalloproteinases (MMP) were analysed by western blot and zymography. KEY RESULTS: ATL-1 inhibited EC adhesion to fibronectin via interaction with its specific receptor. Furthermore, VEGF-induced MMP-9 activity and expression were reduced by pretreatment with ATL-1. Because the transcription factor NF-kappaB has been implicated in VEGF-mediated MMP expression and EC proliferation, we postulated that ATL-1 might modulate the NF-kappaB pathway and, indeed, ATL-1 inhibited NF-kappaB nuclear translocation. Pretreatment of EC with ATL-1 strongly decreased VEGF-dependent phosphorylation of phosphainositide 3-kinase (PI3-K) and extracellular signal-regulated kinase-2 (ERK-2), two signalling kinases involved in EC proliferation. Inhibition of VEGF-induced EC proliferation by ATL-1 was antagonized by sodium orthovanadate, suggesting that this inhibitory activity was mediated by a protein tyrosine phosphatase. This was confirmed by showing that ATL-1 inhibition of VEGF receptor-2 (VEGFR-2) phosphorylation correlates with SHP-1 association with VEGFR-2. CONCLUSIONS AND IMPLICATIONS: The synthetic 15-epi-lipoxin analogue, ATL-1, is a highly potent molecule exerting its effects on multiple steps of the VEGF-induced angiogenesis.

Influence of beverage composition on the results of erosive potential measurement by different measurement techniques.

The influence of beverage composition on the measurement of erosive potential is unclear. The aim of this study was to evaluate whether beverage composition influences the measurement of erosive potential and to evaluate the influence of exposure in small and large volumes. Eleven beverages were included: water (control), 3 alcopops, 2 beers and 5 soft drinks. For each beverage 15 bovine enamel samples were used: 5 for chemical and 10 for profilometric analysis. After exposure to the beverages (63 min) the resulting solutions were analyzed for Ca and inorganic phosphorus (P(i)) content. The samples for optical profilometry were submersed sequentially in 500 ml or in 1 ml of the drinks for 3, 6, 9, 15 and 30 min (total 63 min). For some of the beverages high baseline concentrations of Ca (energy drink) or P(i) (cola drink, cola lemon drink, beer, beer lemon) were found. Some of the beverages showed a good correlation between the chemical methods. Profilometry (both for 1 and 500 ml) showed generally lower enamel losses than the chemical methods. Lower enamel losses were found for the profilometry 1 ml compared to the profilometry 500 ml only for the cola drinks. It can be concluded that the composition of the beverages had a significant effect on the determination of the erosive potential with chemical analyses. Drink composition also influenced the effect of small versus large exposure volumes, indicating the need for standardization of exposure parameters.

Microbial populations of an upflow anaerobic sludge blanket reactor treating wastewater from a gelatin industry.

The microbial populations of an upflow anaerobic sludge blanket reactor, used for treating wastewater from the gelatin industry, were studied by microbiological methods and phase-contrast and electron microscopy. Microscopy examination of the sludge showed a complex mixture of various rod-shaped and coccoid bacterial pluslong filaments and verymobile curved rods. In addition free-living anaerobic ciliates and flagellates were also observed. The trophic group population observed in decreasing order of dominance were hydrolytic and acetogenic at 10(6) and sulfate reducing and methanogenic at 10(5). The rate of methane production in anaerobic granular sludge cultivated in growth medium supplement with formate pressurized with H2:CO2 showed a significant increase in methane yield compared with theseed culture containingthe same substrate and atmosphere of N2:CO2. Similar rates of methane production were observed when the growth medium was supplemented with acetate pressurized either with H2:CO2 or N2:CO2. The number of total anaerobic bacteria at 10(7), fecal coliforms and total coliforms at 10(6), and fecal streptococci at 10(3) is based on colony counts on solid media. The four prevalent species of facultative anaerobic gram-negative bacteria that belong to the family of Enterobacteriaceae were identified as Escherichia coli, Esherichia fergusonii, Klebsiella oxytoca, and Citrobacter freundii. The species Aeromonas hydrophila, Aeromonas veronii, Acinetobacter iwoffi and Stenotrophomonas maltophila were the most frequently isolated glucose fermenting and nonfermenting gram-negative bacilli.

ATL-1, a synthetic analog of lipoxin, modulates endothelial permeability and interaction with tumor cells through a VEGF-dependent mechanism.

Lipoxins (LX) and 15-epi-LX are lipids with a potent inhibitory effect on angiogenesis, in different models in vivo and in vitro. ATL-1, a synthetic analog of 15-epi-LXA4, inhibits various actions stimulated by vascular endothelial growth factor (VEGF). However, LX actions on endothelial cells (EC) in tumor-related contexts are still unknown. Here, we investigated the modulation of EC by ATL-1, in a model that mimics tumor extravasation. We observed that the analog inhibited endothelial permeability induced by VEGF, through the stabilization of VE-cadherin/ß-catenin-dependent adherens junctions. We tested the ability of MV3 cells, a highly metastatic melanoma cell line, to transmigrate across unchallenged EC monolayers for 18 h, as compared to NGM normal melanocytes. ATL-1 was able to inhibit only melanoma extravasation. MV3 cells secrete large amounts of VEGF and we observed that ATL-1 per se did not alter this ability. Melanoma cells skills to crossing endothelial monolayers were due to the steady accumulation of tumor-derived VEGF. When endothelial cells were challenged with exogenous VEGF, added at levels comparable to those secreted by MV3 cells over 18 h, and a short-term (4h) transendothelial migration assay was performed, both melanoma and melanocyte cells were able to extravasate, and ATL-1 was able to block the passage of both cells. These results indicate that ATL-1 has a potent inhibitory effect on the permeability induced by VEGF, and that this pharmacological effect could be used to block tumor extravasation across endothelial barriers, with a possible prospect of reducing the haematogenic spread of cancer cells.

Estimated erosive potential depends on exposure time.

OBJECTIVES: Evaluate erosive potential of beverages, using exposure times from 3 to 30 min, and to analyse the relationship between erosion and several drink parameters. METHODS: pH, calcium, phosphate and fluoride concentration, saturation, titratable-acidity to pH 5.5 and the viscosity of sixteen beverages were measured or calculated. Enamel samples (N = 90) were serially exposed to 1 ml of the beverages for 3, 6, 9, 15 and 30 min and erosion was measured as the loss of calcium to the beverage. Rate of erosion per min was calculated by linear curve fitting using all exposure times. Linear regression analysis was performed to determine the correlation between erosion and the drink parameters. A limited multivariate analysis was performed for the outcome parameter with the highest univariate correlations (erosion per minute) and 4 drink variables. RESULTS: A negative relationship was observed only for pH for all exposure times. Only for erosion per min a significant relationship with pH and saturation was found. In a model for erosion per min using only saturation, fluoride concentration, titratable acidity and viscosity, both saturation and viscosity were shown to have a significant effect (p = 0.01 and p = 0.05, respectively). CONCLUSION: Exposure times between 3 and 30 min result in very different estimates of erosive potential. There is no sound theoretical ground for preferring one or other exposure time/outcome as being more clinically relevant. CLINICAL RELEVANCE: This study shows that effect of the choice of study methodology on the measurement of erosive potential of beverages is large.

Influência das condições térmicas do galpão de espera climatizado na mortalidade pré-abate de frangos de corte / The influence of thermal conditions from an environmentally controlled preslaughter lairage on mortality of broiler chickens

Avaliou-se o efeito das condições ambientais da espera pré-abate na mortalidade de frangos de corte, mantidos em galpão climatizado em um abatedouro comercial. Foram avaliados 215 caminhões de transporte de um abatedouro comercial quanto ao número de aves mortas durante as operações pré-abate e à temperatura retal dos animais em condição de espera no abatedouro. Os dados foram obtidos no galpão de espera climatizado por meio de ventiladores e nebulizadores, em diferentes períodos: manhã, tarde e noite. As variáveis térmicas, distância, tempo de espera e densidade de aves por caixa foram consideradas na análise. Destacaram-se os efeitos da temperatura, da umidade relativa e do tempo de espera na variação tanto do número de aves mortas por caminhão quanto da temperatura retal dos animais. Com relação à interação entre temperatura e umidade relativa interna do galpão de espera, houve eficiência do ambiente climatizado no abatedouro quanto à diminuição do número de aves mortas. A climatização foi mais expressiva na redução do número de aves mortas e da temperatura retal quando a temperatura do galpão de espera estava entre 21 e 24°C, principalmente em condições de umidade relativa interna em torno de 70%. Nessas condições climatizadas, o tempo de espera acima de duas horas proporcionou maior conforto às aves.

The effect of environment conditions of preslaughter lairage on broiler mortality was assessed in the present study. Daily preslaughter data from 215 broiler flocks were recorded regarding the number of dead chickens during preslaughter operations and rectal temperature of broilers submitted to a lairage environment prior to slaughtering. A holding area at a slaughterhouse with environmental control was assessed. The thermal variables, distance between farms and slaughterhouse, lairage time and density of birds per cage was considered in this analysis. The effect of dry-bulb temperature, relative humidity and lairage time on death before arrival and rectal temperature was highlighted in this assessment. Regarding the interaction between temperature and inside relative humidity, a decrease in preslaughter mortality was observed due to exposure to a controlled environment. The environment control promoted an expressive reduction in dead birds and rectal temperature when the temperature inside the holding area was between 21 and 24 °C, mainly with inside relative humidity around 70%. In these environment control conditions lairage above 2 hours was necessary to reduce the thermal stress of birds.

The contribution of extensin network formation to rapid, hydrogen peroxide-mediated increases in grapevine callus wall resistance to fungal lytic enzymes.

Grapevine (Vitis vinifera cv. Touriga) callus cell walls contain a high level of the monomeric extensin, GvP1. Hydrogen peroxide stimulus of these cultures causes the rapid loss of monomeric GvP1, concomitant with marked increases in insoluble GvP1 amino acids and wall resistance to digestion by fungal lytic enzymes. JIM11 immunolocalization studies indicated that monomeric and network GvP1 were evenly distributed in the callus cell wall. These primary cell walls were used to investigate the specific contribution of extensin and other ionically bound cell-wall proteins to hydrogen peroxide-mediated increases in resistance to fungal lytic enzymes. This was performed by removing ionically-bound proteins and assaying for hydrogen peroxide-enhanced resistance after the addition of selected protein fractions. The results indicate that hydrogen peroxide-induced increases in resistance to digestion by fungal lytic enzymes require a co-operative action between network extensin formation and the electrostatic interaction of additional wall proteins with the extracellular matrix.

The regional distribution of neuronal glycogen in the opossum brain, with special reference to hypothalamic systems.

Neurons that accumulate glycogen have been identified in the opossum brain stem and diencephalon by a modified histochemical method using alcoholic solutions and fuchsin proper (pararosanilin) rather than the Schiff reagent (leucosulphite derivative). Several of the glycogen-positive cell groups such as the mesencephalic trigeminal nucleus and the brainstem somatic and special visceral efferent nuclei have been previously detected in the developing brain of small, common laboratory mammals. Scattered glycogen-containing neurons also appear in the dorsal thalamus and basal forebrain. A conspicuous, often Golgi-like accumulation of glycogen has been found in neurons of the magnocellular and parvocellular hypothalamic systems. Together with available data on the metabolic rate of marsupials, our results suggest that the patterns of glycogen deposition may be common to several vertebrates and may be a constant although not exclusive property of cells with axonal endings outside the blood-brain barrier.

Astroglial differentiation in the opossum superior colliculus.

Glial markers, namely, vimentin, glial fibrillary acidic protein (GFAP), and glycogen, as well as accumulation of axon-borne horseradish peroxidase (HRP), were used to visualize radial glial cells in the developing opossum superior colliculus (SC) and to follow changes in young astrocytes of the superficial layers. Vimentin, GFAP, and glycogen are relatively abundant in elements of the median ventricular formation (MVF), which persists at least as late as weaning time, i.e., postconception day 103, postnatal day 90 (PND90). Radial profiles and end-feet in the remaining collicular sectors (main radial system, MRS) are also vimentin-positive but show little or no glycogen or anti-GFAP staining. The numeric density of MRS profiles is very high at the final stages of neuronal migration (PND12) but falls to vestigial numbers by PND 56-60. Antivimentin staining and filling of MRS profiles by axon-borne HRP disappear in parallel. Before total regression of MRS profiles, young astrocytes of the superficial gray layer exhibit a transiently high GFAP expression that is not found in those of the subjacent layers. The results suggest that 1) radial glia at or near the collicular midline are well equipped for a mechanical supportive role, and their abundant glycogen accumulation may reflect their eventual transformation in cells with high glycolytic metabolism, including tanycytes; 2) in most collicular sectors, some radial glia cells persist for long periods after cessation of neuronal migration and may interact with afferent fibers coursing through the superficial neuropil; 3) radially oriented astrocytes of the superficial gray layer exhibit a transiently high GFAP expression that is temporally correlated with late transformations of the retinocollicular projections.