Dynamics of hematological blood parameters of white rats in postnatal ontogenesis

In experiments on Wistar rats, dynamics of blood hematologic and rheological parameters was studied from birth to the 3-month age. Results of this study indicate an intensive activity of the red bone marrow anlagen for the first 3 weeks of postnatal ontogenesis. This is manifested as an increase of the number of cells in the erythron system, a change of the erythrocyte acidic resistance, and a shift in the leukocytic formula. The boundary between the 2nd and 3rd weeks of ontogenesis is a critical period: there occur a sharp deceleration of erythropoiesis and a change of the age-related erythrocyte composition and of the blood leukocyte ratio towards the parameters characteristic of adult rats. The rise in the number of leukocytes and erythrocytes does not stop after this period. Until the 4-week age, the deformability index increases to be accompanied by an increase of hemoglobin hydration and improvement of the erythrocyte toroid shape. Subsequently, inversion of these processes occurs.     

Methyltransferase-inhibition interferes with neuronal differentiation of P19 embryonal carcinoma cells

We have analyzed the importance of substrate methylation by S-adenosylmethionine-dependent methyltransferases for neuronal differentiation of P19 embryonal carcinoma cells. We show that treatment of cells with methyltransferase inhibitor adenosine dialdehyde (AdOx) interferes with neuronal differentiation. Retinoic acid (RA) and AdOx co-treated cells had a decreased number of neurites and a flattened morphology compared with cells differentiated by RA. Also, the amount of neuronal class III tubulin (Tuj1) decreased from 76% to 9.6% with AdOx-treatment. Gene expression levels of wnt-1, brn-2, neuroD, and mash-1 were also down-regulated by AdOx-treatment. But AdOx-treatment did not up-regulate BMP-4 and GFAP genes. Treatment of RA decreased E-cadherin expression during neuronal differentiation. However, in AdOx/RA co-treated cells, E-cadherin expression was restored to the control level. Also, mRNA expression of N-cadherin decreased with AdOx-treatment. Taken together, these data show that methylation reactions might influence the cell-fate decision and neuronal differentiation of P19 cells.     

Effects of Ionizing Radiation and Aluminum Chloride on Protein of Glial Intermediate Filaments in the Rat Brain

We studied the effects of irradiation with X-rays (the total dose of 0.0129 C/kg was attained over 7, 14, or 21 days), increased entry of Al³⁺ into the organism (0.2% AlCl₃ in drinking water), and the combined influence of these factors for 21 days on the contents of the soluble and filamentous forms of glial fibrillary acidic protein (GFAP) in the tissues of the hippocampus, cerebellum, and neocortex of albino rats. After irradiation for 7 days, a clear trend toward drops in the GFAP contents in the structures under study was observed, while irradiation in the same dose, but for 14 or 21 days, resulted in increases in the contents of both GFAP forms (within a range of 13-29%, as compared with the control). Entry of aluminum chloride with water also resulted in an increase in the GFAP contents in all studied structures; changes in the filamentous form were more intensive. The combined influence of irradiation and Al³⁺ resulted in more intensive shifts in the GFAP levels; the content of its filamentous form increased in all structures by about 50%, while shifts of the soluble form were somewhat smaller.     

Sodium nitrite induces acute central nervous system toxicity in guinea pigs exposed to systemic cell-free hemoglobin

Systemic cell-free hemoglobin (Hb) released via hemolysis disrupts vascular homeostasis, in part, through the scavenging of nitric oxide (NO). Sodium nitrite (NaNO₂) therapy can attenuate the hypertensive effects of Hb. However, the chemical reactivity of NaNO₂ with Hb may enhance heme- or iron-mediated toxicities. Here, we investigate the effect of NaNO₂ on the central nervous system (CNS) in guinea pigs exposed to systemic cell-free Hb. Intravascular infusion of NaNO₂, at doses sufficient to alleviate Hb-mediated blood pressure changes, reduced the expression of occludin, but not zona occludens-1 (ZO-1) or claudin-5, in cerebral tight junctions 4 h after Hb infusion. This was accompanied by increased perivascular heme oxygenase-1 expression, neuronal iron deposition, increased astrocyte and microglial activation, and reduced expression of neuron-specific nuclear protein (NeuN). These CNS changes were not observed in animals treated with Hb or NaNO₂ alone. Taken together, these findings suggest that the use of nitrite salts to treat systemic Hb exposure may promote acute CNS toxicity.     

Glial cells positive for glial fibrillary acidic protein in the neurohypophysis of the Djungarian hamster (Phodopus sungorus)

Summary The presence and distribution of the glial fibrillary acidic protein (GFAP; an astrocytic marker protein associated with glial filaments) in the neurohypophysis of the Djungarian hamster (Phodopus sungorus) were investigated immunohistochemically. Our study revealed characteristic GFAP-staining patterns within the median eminence, infundibular stem and neural lobe. In the whole neurohypophysis, few glial cells showed immunoreactivity. In the neural lobe, immunopositive pituicytes appeared preferentially in the periphery. At the ultrastructural level, we found some pituicytes containing filaments, most notably in their processes. We thus demonstrated that, in contrast to the GFAP-immunoreactivity of cultured pituicytes, pituicytic GFAP-expression in vivo coincides with the presence of electron-microscopically detectable filaments.     

Differential Effects of GM1 Ganglioside Treatment on Glial Fibrillary Acidic Protein Content in the Rat Septum and Hippocampus After Partial Interruption of Their Connections

Abstract: The glial fibrillary acidic protein (GFAP) content was investigated using immunoblotting techniques in the septum and hippocampus of the rat after bilateral lateral fimbria transection. Seven days after surgery GFAP content increased significantly both in the septum (140% of control) and hippocampus (120% in dorsal, the less denervated, and 145% in the most denervated ventral part), indicating the occurrence of reactive gliosis. The GM1 treatment caused statistically significant attenuation of GFAP increment in all hippocampal parts. In contrast, GM1 treatment has no influence on the increase of GFAP content in the septum. Results suggest a differential effect of GM1 on the two gliotic reactions formed as a consequence of the lesion at the level of the source of innervation (septum) and the target (hippocampus).     

Utilization and leaching of nitrate from two Deschampsia-dominated heathland sites: a lysimeter study using intact soil columns*

High deposition levels of atmospheric ammonia in the Netherlands have led to the major replacement of dwarf shrubs by grasses and to elevated nitrification rates in acid heathland soils. In order to study the efficacy of a naturally established grass-heath of Deschampsia flexuosa at capturing NO^- ₃, an outdoor 'mesocosm' lysimeter experiment was set up with relatively large and undisturbed soil columns from two Deschampsia-dominated heathland sites. One of the sites (Ede) had a relatively high rate of nitrate production, whereas the other (Hoorneboeg [HB]) showed practically no nitrate formation. For part of the Ede columns, the fate of labeled nitrate, split-applied at two rates (30 or 150 kg ha^-1 yr^-1) during two seasons, was studied. D. flexuosa was highly effective in acquiring fertilizer nitrate, as demonstrated by (1) distinct rises in foliar NRA, especially at high N; (2) increased ¹⁵N enrichments in all plant components with N rate; (3) significant increases in organic-N and carboxylate concentration in several plant compartments; and (4) clear shifts in biomass allocation in favour of the aboveground tissues.After 18 months at low N, an average 39 and 23% of the applied N was immobilized in the plant and soil compartments, respectively; at high N rate, corresponding recoveries were 33 and 20%. Total leaching of nitrate (beyond a depth of 35 cm) from the unfertilized Ede columns corresponded to an annual loss of 1.9 kmol N ha^-1, whereas leaching was virtually zero from HB columns. Relatively high amounts of N leached from the fertilized columns with apparent fertilizer recovery in the leachate reaching an average 60% at high N. However,¹⁵ N analyses revealed only recoveries of 2.0% (low N) and 7.2% (high N) of the applied N in the leachate. From columns where the plant cover had been removed, apparent and real leaching losses reached values of >100 and 10% of the applied N, respectively. Hence, soil-derived N appeared by far the major source of leaching. Unplanted and unfertilized HB columns displayed high rates of nitrification and leached high amounts of nitrate, suggesting a plant-induced repression of the in situ nitrification at this site.On average, planted columns had lost 37% (low N) and 40% (high N) of the applied N, whereas unplanted lysimeters had lost 89% (for both low and high N). The N not recovered was presumed lost by denitrification due to favourable conditions with respect to nitrate concentration, moisture, carbon supply, and temperature.     

Purification and characterization of the central segment of prothymosin-alpha: methodology for handling highly acidic peptides.

Prothymosin-alpha is a highly acidic protein consisting of 110 amino acids. The central segment of this protein, residues 51-89, is thought to be involved in metal binding which may be necessary for its physiological function. To carry out studies of this peptide, this central segment was synthesized in a linear fashion using Fmoc-based methods on rink amide MBHA resin. However, this peptide could not be purified with the typical straightforward approach of RP HPLC followed by negative mode electrospray ionization mass spectrometry (ESI-MS). This was attributed to the high proportion of acidic residues: 26 out of the 39 residues are aspartic and glutamic acids. The acidity of the peptide prevented retention on the RP HPLC column. Additionally, the ability of the highly negatively charged peptide to retain sodium ions prevented molecular weight determination with ESI-MS. A systematic approach to the purification of this highly acidic peptide was undertaken. Ultimately, strong anion exchange chromatography was used to purify the peptide. Extensive desalting using dialysis was required prior to ESI-MS, and the choice of the buffer proved to be critical. In the end, a purification method was devised that yielded a highly purified peptide and is readily compatible with analysis by ESI-MS.     

Deep placement of Mn fertiliser on a sandy soil increased grain yield and reduced split seed in Lupinus angustifolius

Experiments were conducted over two years with Lupinus angustifolius L. on a site with acid sandy soil near Esperance, Western Australia to determine if deep placed manganese fertilizer increases lupin grain yield. Manganese at 4 and 8 kg ha⁻¹ was placed below the surface immediately before sowing at 4, 20 and 30 cm and 4, 8, 12, 16 and 20 cm in 1987 and 1988 respectively. Foliar Mn applied at 1 kg ha⁻¹ when the first order laterals were in mid-flowering stage, was also compared. Increasing the depth of Mn placement increased grain yield in both years. The deepest placed Mn increased grain yields by 255 kg ha⁻¹ (10%) and 430 kg ha⁻¹ (106%) in year 1 and year 2 over the shallow (4 cm) placed Mn. The higher responses to deep placed Mn occurred in year 2, the year with the driest spring and most intense aphid infestations. Foliar applied Mn was as effective as most deep placed Mn treatments, except for the highest rate (8 kg ha⁻¹) at the greatest depth (20 cm) in year 2. The higher rate of applied Mn gave the best grain yields. This revised version was published online in June 2006 with corrections to the Cover Date.