On the staining of elastic fibers with alum hematoxylins.
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A method for the cytophotometric estimation of ribonucleic acid in tissue sections using gallocyanin-chrome alum is described. The dye obeys Beer's law in gelatin sections. The effect of deoxyribonuclease on the staining of ribonucleic acid is also investigated. The results indicate that this method is of value in the quantitation of ribonucleic acid.
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Nondissected ovaries of tuber-bearing Solanum sp. were stained with Mayer's hemalum, a positive stain for chromatin and nucleoli, and then optically cleared with methyl salicylate, a clearing agent. Clarity, resolution and contrast within the ovules dissected from ovaries were comparable to those of sectioned, paraffin embedded ovaries. Contrast within ovules greatly exceeded that of unstained and nonspecifically stained clearings, and eliminated the need of special optics, i.e., Nomarski interference-contrast optics, for optimal viewing and photography. Much less time and labor were required than for embedded specimens. Usefulness of the technique for cytogenetic and cytological research was verified by analyzing meiosis and other features of megasporogenesis and megagametogenesis in normal, and in two meiotic mutants, of Solanum. The results illustrate the usefulness of combined Mayer's hemalum staining and methyl salicylate clearing, and suggest additional stain-clearing agent combinations have potential for cytological and cytogenetic research. Preliminary results with other species suggest the technique may also be useful for classroom instruction.
Two stock solutions are composed as follows: A) aluminum sulfate, sodium iodate and acetic acid in aqueous propylene glycol and B) hematoxylin in pure propylene glycol. When combined in specified proportions the stock solutions yield aluminum-hematein dissolved in nontoxic propylene glycol. The ready-to-use stain, prepared in small volumes as needed, performs well in paraffin sections of plant tissues.
The influence of alum and liposomal phospholipids on interferon-gamma-(IFN-gamma), IFN-gamma/N-acetylmuramyl-L-alanyl-D-isoglutamine- (MDP) or IFN-gamma/tumor necrosis factor-alpha- (IFN-gamma/TNF-alpha) induced macrophage nitric oxide (NO) synthesis has been investigated. IFN-gamma induced NO synthesis in a dose-dependent manner. TNF-alpha and MDP did not induce NO synthesis, but interacted synergistically with sub-optimal doses of IFN-gamma. Alum strongly inhibited IFN-gamma-induced NO synthesis (ID50 25 microgram/ml). Liposomes composed of dipalmitoylphosphatidylcholine (DPPC) had no effect on IFN-gamma-induced NO synthesis. IFN-gamma-induced NO synthesis was stimulated by DPPC/dimyristoylphosphatidylglycerol (DMPG) liposomes (9:1 mol ratio, ED50 45 nmol phospholipid/ml), and inhibited by DPPC/dipalmitoylphosphatidylethanolamine (DPPE) liposomes (9:1 mol ratio, ID50 > 500 nmol phospholipid/ml), and DPPC/phosphatidylserine (PS) liposomes (7:3 mol ratio, ID50 150 nmol phospholipid/ml). Alum, DPPC/PE and DPPC/PS liposomes also inhibited IFN-gamma/MDP- and IFN-gamma/TNF-alpha-induced NO synthesis. Neither alum or the liposome preparations had significant toxicity towards macrophages in vitro at concentrations that induced maximal inhibition or stimulation of IFN-gamma-induced NO synthesis. Immunization of mice with alum-adsorbed and liposome-incorporated bovine serum albumin (BSA) demonstrated that enhancement or reduction of both IgG antibody and the proportion of IgG2a/IgG2b was correlated with stimulation or inhibition of IFN-gamma-induced NO synthesis.
Development of cost-effective amendments for treating dairy slurry has become a critical problem as the number of cows on farms continues to grow and the acreage available for manure spreading continues to shrink. To determine effectiveness and optimal rates of addition of either alum or zeolite to dairy slurry, we measured ammonia emissions and resulting chemical changes in the slurry in response to the addition of amendments at 0.4, 1.0, 2.5, and 6.25% by weight. Ammonia volatilization over 96 h was measured with six small wind tunnels with gas scrubbing bottles at the inlets and outlets. Manure samples from the start and end of trials were analyzed for total nitrogen and phosphorus, and were extracted with 0.01 M CaCl2, 1.0 M KCl, and water with the extracts analyzed for ammonium nitrogen, phosphorous, aluminum, and pH. The addition of 6.25% zeolite or 2.5% alum to dairy slurry reduced ammonia emissions by nearly 50 and 60%, respectively. Alum treatment retained ammonia by reducing the slurry pH to 5 or less. In contrast, zeolite, being a cation exchange medium, adsorbed ammonium and reduced dissolved ammonia gas. In addition, alum essentially eliminated soluble phosphorous. Zeolite also reduced soluble phosphorous by over half, but the mechanism for this reduction is unclear. Alum must be carefully added to slurry to avoid effervescence and excess additions, which can increase soluble aluminum in the slurry. The use of alum or zeolites as on-farm amendment to dairy slurry offers the potential for reducing ammonia emissions and soluble phosphorus in dairy slurry.
Groups of BALB/c mice were sham infected or inoculated intranasally (IN) with live RSV. From Day 4 to 8 after infection, the animals were exposed IN to ovalbumin (OVA) with or without alum adjuvant. At different intervals, levels of OVA concentration in serum, IgG-anti-OVA antibody activity in serum, and IgA-anti-OVA antibody activity in bronchial washings were determined, employing the ELISA technique. IgE-anti-OVA antibody titers in serum and bronchial washings were assessed by PCA. OVA concentrations in serum were significantly higher in RSV-infected animals compared to uninfected controls. The use of alum adjuvant also increased OVA uptake in uninfected animals but to a lesser extent than RSV infection. RSV-infected animals developed significantly higher OVA-specific antibody titers of IgG isotype in serum and IgA isotype in bronchial washings than the uninfected controls, while alum enhanced the immune response less markedly but still significantly in uninfected mice. An IgE antibody response to OVA in serum was demonstrable in 50% of RSV-infected mice immunized IN with OVA and alum, while all uninfected animals and RSV-infected animals immunized with OVA alone (without adjuvant) failed to develop a detectable IgE response. These findings suggest that infections with viral agents such as RSV may function as adjuvants for other antigens inhaled during acute respiratory infection. These observations may explain the alterations in the immune response to other antigens in patients with acute viral-induced bronchopulmonary diseases.
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Disturbances in metal-ion transport, homeostasis, overload and metal ion-mediated catalysis are implicated in neurodegenerative conditions such as Alzheimer's disease (AD). The mechanisms of metal-ion induced disruption of genetic function, termed genotoxicity, are not well understood. In these experiments we examined the effects of non-apoptotic concentrations of magnesium-, iron- and aluminum-sulfate on gene expression patterns in untransformed human neural (HN) cells in primary culture using high density DNA array profiling and Western immunoassay. Two week old HN cells were exposed to low micromolar magnesium, iron, or aluminum for 7 days, representing trace metal exposure over one-third of their lifespan. While total RNA yield and abundance were not significantly altered, both iron and aluminum were found to induce HSP27, COX-2, betaAPP and DAXX gene expression. Similarly up-regulated gene expression for these stress-sensing, pro-inflammatory and pro-apoptotic elements have been observed in AD brain. The combination of iron and aluminum together was found to be particularly effective in up-regulating these genes, and was preceded by the evolution of reactive oxygen intermediates as measured by 2',7'-dichlorofluorescein diacetate assay. These data indicate that physiologically relevant amounts of iron and aluminum are capable of inducing Fenton chemistry-triggered gene expression programs that may support downstream pathogenic responses and brain cell dysfunction.
Changes in laying-hen performance due to the effect of graded levels of dietary fluoride (F) have been documented earlier. The present study examined the effect of dietary F and aluminum (Al) on laying hen performance, and F levels in plasma, soft tissues, muscle, bone, and eggs. Two hundred and seventy Single Comb White Leghorn (SCWL) layers were fed a basal diet containing the following F and Al levels: control; 100 ppm F; 80 ppm Al; 100 ppm F/80 ppm Al; 100 ppm F/40 ppm Al; 1300 ppm F; 1040 ppm Al; 1300 ppm F/1040 ppm Al, and 1300 ppm F/520 ppm Al. After 112 days of feeding, the blood, tissue, bone, and egg samples were collected for analysis. High F intake significantly depressed feed intake and hen-day production and elevated serum, tissue, bone, and egg F levels. Dietary Al minimized the effect of high fluoride feeding. The mean fluoride concentration for low (80 ppm Al diet) vs. high F (1300 ppm) intake was, respectively: plasma, .2 vs. 10.1 ppm; liver, 4.5 vs. 19.2 ppm; kidney, 2.9 vs. 31.8 ppm; pectoralis muscle, 3.6 vs. 6.7; tibia muscle homogenate, 3.9 vs. 21.1 ppm, and bone, 611 vs. 2600 ppm. The majority of the F transferred to the egg was incorporated into the shell (2.8 to 307.1 ppm). Low F levels were found in albumen (fresh, .4 to 1.0 ppm) and yolk (dry fat free, 3.1 to 18.4 ppm). The data suggested that dietary F is readily accumulated in bone and kidney; however, liver and muscle are less responsive.(ABSTRACT TRUNCATED AT 250 WORDS)
The effect of dietary aluminum sulfate on Ca and P metabolism was studied using 1-day-old male broiler chicks. In Experiment 1, practical diets providing .90% Ca plus .45% available P (Pav), .90% Ca plus .78% Pav, 1.80% Ca plus .45% Pav, or 1.80% Ca plus .90% Pav were fed with 0 or .392% A1 as aluminum sulfate for 21 days. The control diet (.90% Ca plus .45% Pav) without added A1 was fed to all chicks during Days 22 to 49. In general, A1 significantly (P less than .05) decreased BW gain, feed intake, gain:feed ratio, plasma inorganic P (Pi), tibia breaking strength, tibia weight, percentage of tibia ash, and plasma Zn, measured at Day 21. Elevating Pav increased BW gain, feed intake, gain:feed ratio, tibia weight and plasma Zn, and decreased plasma total Ca in the presence of .392% A1 plus 1.80% Ca. Plasma Pi, tibia breaking strength, and percentage of tibia ash were increased by raising dietary Pav in the presence of .392% A1 with either level of Ca. Negative effects of dietary A1 on feed intake and BW persisted through Day 49. In Experiment 2, a control diet (.90% Ca, .45% Pav) was fed for ad libitum access either alone or supplemented with .2% A1 as aluminum sulfate or with an equivalent amount of sulfate provided by potassium sulfate. The control diet was also pair-fed to chicks given .2% A1. Dietary A1 significantly depressed weight gain, feed intake, gain:feed ratio, and plasma Pi.(ABSTRACT TRUNCATED AT 250 WORDS)
Experiments were conducted to determine if the high ion-exchange capacity of sodium aluminosilicate (ZA) marketed as ETHACAL Feed Component or its aluminum content or both are related to the beneficial effect of ZA on egg specific gravity (ESG). In Experiments 1 and 2, ZA was compared with hydroxy-sodalite (HS) and carnegieite, which have the same chemical formula as ZA but little or no ion-exchange capacity. Two levels of ZA (0 and 1.5% of the diet) and three levels of HS (0, .75, and 1.5%) were fed in Experiment 1. In Experiment 2, 0, .75, and 1.5% of ZA and carnegieite were fed. In Experiment 3, ZA, carnegieite, aluminum sulfate, and aluminum phosphate were fed at levels calculated to contain .148 and .101% aluminum. Criteria evaluated were ESG, egg production, feed consumption, and egg weight. Egg specific gravity was significantly improved by ZA in all experiments and was not affected by HS, carnegieite (Experiments 1 and 2), aluminum sulfate (.148% Al), or aluminum phosphate (Experiment 3). Carnegieite and aluminum (.101%) from aluminum sulfate increased ESG in Experiment 3. In Experiments 1 and 3, egg production and feed consumption were not influenced by various treatments. Carnegieite and ZA reduced egg production and feed consumption in Experiment 2. It was concluded that the ion-exchange capacity and aluminum content of ZA may contribute to its beneficial effect on ESG.
Two experiments were conducted to characterize the toxicity and evaluate the efficacy of alum to increase intestinal strength in young broiler chicks. Cobb x Cobb male broiler chicks were placed in an experimental design consisting of six dietary treatments of alum (control, 0.23, 0.47, 0.93, 1.9, and 3.7%) with four replicate pens of 10 broilers per pen. The chicks were housed in electrically heated batteries and provided the treatments for ad libitum consumption from 1 d to 3 wk of age. Alum significantly (P < or = 0.05) decreased body weights at 1.9 and 3.7% in Experiment 1 and at 0.93, 1.9, and 3.7% in Experiment 2. Feed conversion and the relative weight of the gizzard were increased in both experiments at 3.7%. Serum phosphorus was decreased at 1.9 and 3.7% in Experiment 1 and at 3.7% in Experiment 2. Intestinal and bone strength were decreased in both experiments at 3.7%. Bone ash was reduced at 3.7% in Experiment 2, bone S levels increased at 1.9 and 3.7% in Experiment 1 and at 3.7% in Experiment 2, and bone Al levels were elevated in both experiments at 3.7%. Muscle levels of P and S decreased, and that of Ca increased at 3.7%. Aluminum levels were not elevated in muscle tissues. These data indicate that alum can be toxic to young broiler chicks, but at levels that would not be expected to be reached through litter consumption, and that alum did not increase intestinal strength.
AIM: To study the therapeutic effectiveness of intra-tumor injection of Xiao-Zhi-Ling(XZL) on transplanted hepatoma in rats. METHODS: Sixty rats were divided into 3 groups (groups S, X and E), 20 in each. Different drugs were injected into the implanted hepatoma (Group S with 0.2 ml saline as control, group X with 0.2 ml XZL, group E with 0.2 ml ethanol). After 3 days and 8 days respectively, we detected the hepatoma volume (HV), the level of albumin, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) in serum, and the expression of proliferating cell nuclear antigen (PCNA) in hepatoma. RESULTS: The results were obtained after 3 days, the HVs in groups X and E were smaller than those in group S (group X vs S P=0.010(*), group E vs S P=0.002(*), P<0.05). The levels of ALT and AST in group S and X were lower than those in group E (ALT Group S vs E P=0.019(*), group X vs E P=0.003(*), P<0.05; AST group X vs E P=0.002(*), P<0.05). The levels of ALP and PCNA labeling index in group X were lower than those in group S and E (ALP group X vs S P=0.000(*), group X vs E P=0.000(*). P<0.05; PCNA group X vs S P=0.008(*), group X vs E P=0.048(*), P<0.05). The levels of creatinine in group S were lower than those in group E (group S vs E P=0.017, P<0.05). The degree of tumor necrosis in group S was lower than those in groups X and E (group S vs X P=0.006(*), group S vs E P=0.006(*), P<0.05). After 8 days, the HVs in groups X and E were smaller than those in group S (group X vs S P=0.007(*), group E vs S P=0.004(*), P<0.05). The difference of HVs between groups X and E was not significant. The levels of albumin, ALT, AST and creatinine in group X were not higher than those in other groups, the levels of ALP and PCNA in group X were lower than those in groups S and E (ALP group X vs E P=0.006(*) P<0.05; PCNA group X vs S P=0.044(*), group X vs E P=0.021(*), P<0.05). The degree of tumor necrosis in group S was lower than that in groups X and E (group S vs X P=0.001(*), group S vs E P=0.002(*), P<0.05). CONCLUSION: The therapeutic effectiveness of intra-tumor injection of XZL and ethanol on implanted hepatoma is obvious, but the toxicity of XZL on liver function is markedly lower than that of group E, at the same time XZL can inhibit the growth of tumor. XZL is relatively better and safer than ethanol in intra-tumor injection therapy.
Enhancement of the adhesiveness of dental restoratives, such as composite resins, has been clinically essential. A key point in enhancing adhesion is how to treat the dentin surface before bonding. We recently attempted to use an inorganic and non-acidic salt solution for pretreatment to dentin. We paid attention to the salting-in effect of rhodanides which promotes swelling of collagen and gelatin. For pretreatment, we used potassium thiocyanate (KSCN) solution. This study morphologically evaluated the effect of treatment of dentinal smear layer with rhodanides in combination with cross-linking agents such as ferric chloride. Scanning electron microscopy revealed effective removal of the smear layer on the treated dentin surface and obturation of dentinal plugs. Addition of the cross-linking agents to KSCN solution was more effective for pretreatment.
Cr2-/- mice have an impairment in humoral immunity, as shown by the decrease in the Ab titers against T cell-dependent Ags and abnormalities in germinal center formation. Germinal centers are present, but they are decreased in size and number, indicating problems in their development. In this study, we investigated whether this abnormality in germinal center development is associated with problems in the establishment of optimal affinity maturation and the generation of memory B cells, processes closely related to the germinal center reaction. We immunized the Cr2-/- animals with different Ags with or without adjuvants. We showed that, when immunized without adjuvants, complement receptors are absolutely required for optimal affinity maturation. Although limited affinity maturation is elicited in the Cr2-/- Ab response, it is decreased as compared with normal animals. Memory B cell generation is also impaired. In the presence of adjuvants, germinal center development in the Cr2-/- mice is still abnormal, as demonstrated by their decreased size and number. Surprisingly, adjuvants establish optimal affinity maturation and partially restore the amount of Ab produced during the primary response and memory B cell generation. However, adjuvants cannot improve the ability of follicular dendritic cells to retain Ags in the form of immune complexes. These observations indicate that immunization with inflammatory Ags offset some of the immunological abnormalities found in the Cr2-/- mice and show that optimal affinity maturation in the Cr2-/- mice can be achieved in the absence of normal germinal centers.
Control of IgE Ab production is important for the prevention of IgE-related diseases. However, in contrast to the existing information on the induction of IgE production, little is known about the regulation of the production of this isotype, with the exception of the well-documented mechanism involving T cell subsets and their cytokine products. In this study, we demonstrate an alternative approach to interfere with the production of IgE, independent of the activity of T cells, which was discovered during the course of an investigation intended to clarify the mechanism of IgE-selective unresponsiveness induced by surface-coupled liposomal Ags. Immunization of mice with OVA-liposome conjugates induced IgE-selective unresponsiveness without apparent Th1 polarization. Neither IL-12, IL-10, nor CD8(+) T cells participated in the regulation. Furthermore, CD4(+) T cells of mice immunized with OVA-liposome were capable of inducing Ag-specific IgE synthesis in athymic nude mice immunized with alum-adsorbed OVA. In contrast, immunization of the recipient mice with OVA-liposome did not induce anti-OVA IgE production, even when CD4(+) T cells of mice immunized with alum-adsorbed OVA were transferred. In the secondary immune response, OVA-liposome enhanced anti-OVA IgG Ab production, but it did not enhance ongoing IgE production, suggesting that the IgE-selective unresponsiveness induced by the liposomal Ag involved direct effects on IgE, but not IgG switching in vivo. These results suggest the existence of an alternative mechanism not involving T cells in the regulation of IgE synthesis.