Amount Of Reaction Product Research Articles

Blood-brain barrier Ca2+-ATPase cytochemistry: incubation media and fixation methods for differentiating Ca2+-specific ATPase from ecto-ATPase

Ca2+-ATPase cytochemistry frequently uses the incubation medium of Ando et al. that was introduced in 1981. Some studies, however, have suggested that this medium localizes ecto-ATPase in addition to Ca2+-ATPase and that Ca2+-ATPase is sensitive to fixation. Strong activity of the enzyme on the luminal surface of the blood-brain barrier (BBB) also is considered indicative of immature or pathological microvessels. We address here five questions. 1) Is the incubation medium of Ando et al. specific for BBB Ca2+-ATPase or does it also localize ecto-ATPase? 2) How are the two enzymes distributed in the BBB? 3) How would data interpretation be prone to error if the cytochemical study does not use controls identifying ecto-ATPase? 4) Does the amount of reaction product of both enzymes vary significantly when the cortical tissue is exposed to different fixatives? 5) Does the presence of Ca2+-ATPase on the luminal membrane of the BBB necessarily indicate immature or abnormal brain endothelial cells? Adult male Sprague-Dawley rats were perfused with one of two different fixatives and vibratome slices of the brain cortex were incubated in the medium of Ando et al. The controls used were those demonstrating the ecto-ATPase and those that do not. The results indicate that the incubation medium is not specific for Ca2+-ATPase, because it also localizes the ecto-ATPase. Ca2+-ATPase appears to be localized primarily on the luminal surface of the BBB, while ecto-ATPase is localized on both the luminal and abluminal surfaces. The portion of the reaction product contributed by Ca2+-ATPase would not have been identified if the controls uniquely identifying the ecto-ATPase had not been used. The amount of reaction product formed by Ca2+-ATPase is strongly dependent on the type of fixative used. The strong localization of Ca2+-ATPase on the luminal surface of the BBB is not only normal, but also better accounts for the physiological homeostasis of Ca2+ across the blood-brain interface and should not be interpreted as indicative of immature or pathological microvessels.

Genotypical features in the expression of allozymes of β-specific hydrolase of carboxylic esters in wild-type Drosophila melanogaster

The expression level of electrophoretically separated S- and F-allozymes of β-specific esterase (EC 3.1.1.2) in genotypes of wild-type Drosophila melanogaster (males and females) that are monozygous or heterozygous with respect to the locus β-Est is determined by means of computerized densitometry; α-naphthylacetate, β-naphthylacetate, and α-naphthylpropionate are used as the substrates. The intensity of the expression of the esterase is judged from the quantity of reaction product created as a result of simultaneous azo coupling between naphthol and diazonium in 4, 24, 44, and 64 min incubation times. Reliable differences in the expressions of the S- and F-allozymes as a function of the structure of the β-Est locus of genotypically distinct individuals are established. In all the variant experiments, a higher level of summary activity of the S- and F-allozymes of the β-esterase of the heterozygotes by comparison with the individual activity of the F-and S-allozymes of the corresponding homozygotes was demonstrated, independently of the sex of the Drosophila individual. A comparative estimate of the temporal dynamics of the expression of in vitro allozymes of the dominant homozygotes (β-EstS/β-EstS), heterozygotes (β-EstS/β-EstF), and recessive homozygotes (β-EstF/β-EstF) is performed. Possible mechanisms for the occurrence of heterosis according to the character of expression of S- and F-allozymes of β-esterase on the basis of the theory of biochemical enrichment of heterozygote genotypes are considered.

Differential O-3/O-4 regioselectivity in the glycosylation of α and β anomers of 6-O-substituted N-dimethylmaleoyl-protected d-glucosamine acceptors

An assessment of the relative O-3/O-4 reactivities of both methyl α- and β- d-glycosides of N-dimethylmaleoyl (DMM) d-glucosamine acceptors protected at O-6 with benzoyl (Bz), benzyl (Bn), and tert-butyldiphenylsilyl (TBDPS) groups is presented using per-O-benzoylated β- d-galactofuranosyl and per-O-acetylated α- d-galactopyranosyl trichloroacetimidates as glycosyl donors. Using the former donor, the α anomer of the 6-O-benzoylated compound gave exclusive substitution at O-3, whereas the other two compounds with α-configuration kept this site as preferential. The β anomer of the 6-O-benzoylated compound gave the same amounts of reaction products on O-3 and O-4, whereas the other β analogs carried a more reactive O-4. The same reactions were carried out using as donor the less-reactive per-O-acetylated α- d-galactopyranosyl trichloroacetimidate. Although the same trend was found to occur, the O-4 was always relatively more reactive with the pyranosyl donor than with the furanosyl donor, when keeping the remaining factors constant. Furthermore, the β anomers of the acceptor gave almost exclusive substitution at O-4. These observations confirm and extend the utility of these ‘matching’ donor and acceptor reactivities.

Formation and emissions of carbonyls during and following gas-phase ozonation of indoor materials

Ozone concentrations that are several orders of magnitude greater than typical urban ambient concentrations are necessary for gas-phase ozonation of buildings, either for deodorization or for disinfection of biological agents. However, there is currently no published literature on the interaction of building materials and ozone under such extreme conditions. It would be useful to understand, for example in the case of building re-occupation planning, what types and amounts of reaction products may form and persist in a building after ozonation. In this study, 24 materials were exposed to ozone at concentrations of 1000 ppm in the inlet stream of experimental chambers. Fifteen target carbonyls were selected and measured as building ozonation by-products (BOBPs). During the 36 h that include the 16 h ozonation and 20 h persistence phase, the total BOBP mass released from flooring and wall coverings ranged from 1 to 20 mg m −2, with most of the carbonyls being of lower molecular weight (C 1–C 4). In contrast, total BOBP mass released from wood-based products ranged from 20 to 100 mg m −2, with a greater fraction of the BOBPs being heavier carbonyls (C 5–C 9). The total BOBP mass released during an ozonation event is a function of both the total surface area of the material and the BOBP emission rate per unit area of material. Ceiling tile, carpet, office partition, and gypsum wallboard with flat latex paint often have large surface areas in commercial buildings and these same materials exhibited relatively high BOBP releases. The greatest overall BOBP mass releases were observed for three materials that building occupants might have significant contact with: paper, office partition, and medium density fiberboard, e.g., often used in office furniture. These materials also exhibited extended BOBP persistence following ozonation; some BOBPs (e.g., nonanal) persist for months or more at emission rates large enough to result in indoor concentrations that exceed their odor threshold.

Mechanism of the NO2 Conversion to NO2− in an Alkaline Solution

The reaction of NO2 and NaOH aqueous solution at room temperature was studied for elucidating the behavior of gaseous NO2 in an alkaline solution. Experimental runs related to NO2 absorption have been carried out in various pH solutions. The nitrite and nitrate ions formed in these absorption solutions were quantitatively analyzed. In the case of pH 5-12, both of the nitrite and nitrate ions were formed simultaneously. On the other hand, only the nitrite ion was formed when the pH of the absorption solution was higher than 13. In this paper, a new reaction mechanism was proposed to explain the selective formation of nitrite ion in the 10 M alkaline solution. In order to confirm the new reaction mechanism, H2(18)O was used as part of the absorption solution for detecting oxygen gas production. The amounts of reaction products: (18)O(18)O, (18)O(16)O and (16)O(16)O, were quantitatively determined. It was confirmed that the new reaction proceeds mainly in the 10 M alkaline solution.

ToF‐SIMS analysis of model compounds of friction modifier adsorbed onto friction surfaces of ferrous materials

AbstractThe adsorption of model compounds of friction modifiers on ferrous materials has been investigated by time‐of‐flight secondary ion mass spectrometry (ToF‐SIMS). Eleven kinds of model compounds, including carboxylic acids, alcohols, amines and esters, were subjected to lubrication tests. Six compounds among these—carboxylic acids and some amines and esters—showed good lubricity. The ToF‐SIMS spectra of the friction surfaces after being washed with hexane showed that carboxylic acids formed Fe‐carboxylates by reacting with the ferrous material, and esters formed both Fe‐carboxylates and Fe‐alcoholates by reactions of hydrolysates of esters and the ferrous material. For amines, no reaction products were detected but quasimolecular ions, suggesting physical adsorption, were detected from the friction surfaces. For alcohols, no adsorption or reaction was detected from the friction surface. However, clear correlations were not found between the amounts of reaction products or adsorbed materials and lubrication properties such as friction coefficient and width of wear scar. Copyright © 2001 John Wiley & Sons, Ltd.

Platelet-activating Factor (PAF)-dependent Transacetylase and Its Relationship with PAF Acetylhydrolases

Platelet-activating Factor (PAF)-dependent Transacetylase and Its Relationship with PAF Acetylhydrolases

The method for analyzing anhydride formed in poly(butylene terephthalate) (PBT) during thermal and photo-degradation processes and applications for evaluation of the extent of degradation

The pyrolysis GC/MS in the presence of tetramethyl ammonium hydroxide (TMAH) was used to detect anhydride formed in photo-oxidation of poly(butylene terephthalate) (PBT). It was decided that to react anhydride with TMAH to give methyl 4-methoxybutyrate (reaction product). It was confirmed that anhydride was also formed likewise in the thermal degradation. In thermal degradation from 140 to 200°C the amount of reaction product was in agreement with that of COOH end-groups. In view of the kinetics of reaction product and COOH end-groups, thermo-oxidation was dominant from 140 to 180°C, and thermal decomposition dominant at 200°C. On parts (locks) subjected to load in thermal degradation and outdoor exposed mold connectors, the relationships between the breakage rate due to decrease of mechanical strength and the concentration profile depthwise of reaction product was investigated. The decrease of mechanical strength depended on oxidation and decomposition across the entire depth of sample in thermal degradation, while only on the intensity of oxidation in the most superficial 15 μm layer in photo degradation. This method has practical applications for the evaluation of thermo-oxidation and photo-oxidation of PBT.

Interfacial reactions in aluminum/SiC fibre composite electric power cable using low oxygen SiC fibre reinforcement

We have evaluated the interfacial reactions of SiC fibre reinforced Al electrical power cable using low oxygen SiC fibre (Si : 62.4, C : 37.1, 0 : 0.5 mass%), and determined the relationship between the tensile strength and the amount of reaction products at the interface. The following are occurring at the SiC/Al interface: i) diffusion of Al atoms into the SiC fibre, ii) formation of needle–shape Al4C3 compounds, and iii) formation of Al9Si compounds. Formation of Al4C3 and Al9Si compounds at the interface causes the strength of SiC/Al composite electric power cable to deteriorate.

The capillary of the organum vasculosum laminae terminalis (OVLT) in rabbits is more permeable to horseradish peroxidase (HRP) than that in rats.

The distribution of the tracer, horseradish peroxidase (HRP), in the organum vasculosum laminae terminalis (OVLT) of rabbits and rats was examined by electron microscopy. In rabbits, the HRP-diaminobenzidine reaction products were heavily distributed in the OVLT and surrounding brain tissues 10 and 60 min after the injection of HRP (50 mg kg(-1), i.v.), and were retained in the parenchymal tissues at 24 h post-injection. The majority was found in numerous large phagosomes of macrophages located in the perivascular spaces of the vascular beds and in ependymal cells (tanycytes) in the parenchyma. A large amount of reaction product was also localized in the intercellular clefts of the parenchyma. HRP-incorporation was seen in both nerve cells and ependymal cells in the OVLT at 10 min post-injection, but only in nerve cells in the preoptic area at 60 min post-injection. In rats, however, a small amount of the reaction products was observed in the OVLT 10 min after the injection of HRP (50 and 70 mg kg(-1), i.v.), and the levels were markedly reduced at 60 min post-injection. No HRP-incorporation by nerve cells was seen. From these findings, we concluded that the capillary of the OVLT of the rabbit is more permeable to HRP than that of the rat.

Reactivity in the chromium oxide-calcium fluoride system: An empirical approach

The reaction mechanisms observed when sintering loose Cr2O3–CaF2 powder mixtures were analyzed, and the influence of the sintering parameters on the reaction behavior is presented. Using x-ray diffraction (XRD), energy-dispersive spectroscopy (EDS), and differential thermal analysis (DTA) measurements, CaCrO4 was shown to be the reaction product when sintering in air. The reaction occurs in two steps: CaF2 transforms to CaO at the Cr2O3–CaF2 interface, followed by a CaO–Cr2O3 interaction, which creates the reaction product. Scanning electron microscopy (SEM) and x-ray fluorescence (XRF) analysis showed an increasing loss of CaF2 with increasing sintering temperature and heating rate, while an opposite evolution of the amount of reaction product was observed.

Stability of enzymatic indicators of metabolic and neuronal activity in post-ganglionic neurons supplying the urinary tract of aged rats

Aged post-ganglionic neurons supplying the urinary tract exhibit stability in their metabolic and functional status, as demonstrated by quantification of enzyme histochemical reaction products. The major pelvic and thirteenth thoracic (T13) ganglia from 4- and 24-month-old male Wistar rats were incubated under standardized conditions to demonstrate succinate dehydrogenase, NADH-tetrazolium reductase and cytochrome oxidase activities. The amount of reaction product for each enzyme, localized in neuronal perikarya throughout the ganglia, was quantified by microdensitometry. Comparison of mean totals demonstrated stability of all three enzymes with age in both the major pelvic and T13 ganglia and in target organs. Also, the distribution and range of reaction product densities were similar. In both ganglia, the number of NADH-tetrazolium reductase-stained neurons per unit area was reduced in the aged animals. Additionally, nicotinic (DMPP) and muscarinic (McN-A-343) agonists were used to mimic the effects of preganglionic stimulation in the major pelvic ganglia before performing enzyme histochemical reaction. Quantification of these activities demonstrated that the response of aged neurons to preganglionic stimulation was unchanged. These results indicate an overall metabolic stability in post-ganglionic autonomic neurons innervating the urinary tract and in their targets in aged rats. © 1998 Chapman & Hall

Various reactions of sevoflurane with the individual components of soda lime

The various components of commercial soda lime (sodium hydroxide, potassium hydroxide, calcium hydroxide, barium hydroxide) were studied in terms of their reactivity with sevoflurane at its boiling point (59 degrees C). A simple closed system, a reflux cooler, served as a model. Analyses were performed by GC/MS. Besides sevoflurane, we identified four compounds: A, B, C, and D. Free methanol, formaldehyde and formic acid could not be found. Presumably methanol is transferred from an intermediate formalin-semiacetal of the hexafluorisopropanol. Calcium hydroxide and barium hydroxide showed little reaction with sevoflurane, whereas larger amounts of reaction products were observed with sodium hydroxide and potassium hydroxide. The alkali hydroxides of sodalime are presumably responsible for its reaction with halogenated inhalation anaesthetics. We therefore conclude that decomposing reactions of halogenated inhalation anesthetics with dry soda lime could be prevented by using a newly developed soda lime.

Development of hydrogen peroxide (H 2O 2)-generating oxidases in chick organs with special reference to kidneys

The developmental pattern of H2O2-producing oxidases (OX) was studied in chick kidneys (mesonephros, metanephros), intestine, liver, yolk sac and adrenal glands between embryonic days (ED) 5-20 as well as in chick organs after hatching. Sections from snap frozen tissue fixed in cold cacodylate-buffered 2% glutaraldehyde were processed by cerium-DAB-Co-H2O2 methods for benzylamine OX, diamine OX, histamine OX, alpha-hydroxyacid OX, D-amino acid OX (AAOX) and monoamine OX (MAOX). Prenatally, only activities of AAOX and MAOX could be demonstrated. AAOX appeared primarily in the proximal tubular cells of both types of kidneys. In the metanephros the enzyme was also detected in the thick ascending limbs of Henle's loops. The amount of reaction product in tubular cells increased with their maturation. MAOX activity was detected in immature enterocytes, in smooth muscle cells of large systemic arteries (on ED 5-6) as well as in proximal tubular cells of the mesonephros and adrenal gland. Later the enzyme appeared also in smooth muscle cells of the intestinal wall and in endothelial and smooth muscle cells of arterioles of the mesonephros. In the metanephros MAOX was detected at the same locations with a time delay because of a developmental shift of the kidney. Inhibition tests revealed that MAOX differs in epithelial cells from that in smooth muscle cells. Benzylamine OX, diamine OX and histamine OX were detected postnatally in smooth muscle cells of the arterial media and muscularis externa of the intestinal wall with low activities. It is concluded that MAOX and AAOX activities represent useful markers in the development of renal tubules. In addition, MAOX activity can be considered an indicator of maturation of components of the vascular wall.

Reactions of gases on calcareous stones under dry conditions in field and laboratory studies

Dry deposition of gases plays an important role for the deterioration of stone materials and a better understanding of the processes involved will improve our ability to maintain stone monuments and buildings. As a part of an EU-project an investigation with four calcareous stone types have been exposed outdoor at two test sites in Norway for two years. The exposure has been carried out in sheltered position and the amount of reaction products and the penetration depth of SO2 into the stones was determined as soluble sulphate after half a year and after one and two years. Even if most of the sulphate was found in the upper 0.3 mm of the stone, there was an increase in the sulphate content in stone even down to the center of the stone sample. In laboratory tests with SO2, NO2 and changing relative humidity the synergistic effect of NO2 and the importance of the relative humidity was investigated. The uptake rates were calculated from the laboratory studies by analyzing the gas concentrations before and after the exposure chamber. By calculating the deposition velocity from the field study by using the amount of sulphate found in the stones together with the average outdoor concentration of SO2 at the test sites, the values were a magnitude higher than in the laboratory test, highest at the industrial paper mill sites with high concentrations both of SO2 and some hypochlorite and lower in urban atmosphere with fairly low values of SO2 and high values of NO2.

A light microscopical, ultrastructural and immunohistochemical study of spindle-cell adrenocortical tumours of ferrets

Twelve adrenocortical tumours with a variable spindle-cell component in ferrets (six spayed females, three intact females, two castrated males, and one intact male) were examined by light microscopy. One tumour with a moderate spindle-cell component was examined ultrastructurally, and three tumours were studied immunohistochemically. Light microscopy revealed a spindle-cell component in the tumours that varied from a few such cells occupying the stroma between packets of adrenocortical cells to cells in such large numbers that they formed almost the entire substance of the tumour. By light microscopy these spindle cells resembled smooth muscle cells, and the ultrastructural findings, particularly the presence of thin contractile filaments, suggested that the spindle cells were of smooth muscle origin. Immunohistochemical staining revealed that the spindle cells were negative for cytokeratins and S-100 protein but positive for smooth muscle actin. Desmin was readily demonstrated in two tumours but not in the other examined. Vimentin was variable, generally producing a small to moderate amount of reaction product.

Comparative methodological investigations on the cytochemical localization of calcium in brain and inner ear of cichlid fish

Four different methods for calcium precipitation are compared in the optic tectum and the inner ear of the cichid fish, Oreochromis mossambicus. Several parameters are investigated concerning their influences on the reaction product. Three procedures (bichromate, fluoride, and oxalate-pyroantimonate) produce fine-grained deposits, often flocculent in the latter method. The fourth method (potassium-pyroantimonate) generates predominantly coarse-grained reaction product. The calcium content of the deposits is always proven with energy-filtering transmission electron microscopy (EFTEM). In both tissues fine-grained reaction product is found in endoplasmic reticulum and synaptic vesicles, and in addition in some mitochondria and at the cytoskeleton. The coarse-grained deposits of the potassium-pyroantimonate method have a more unspecific distribution. This is the only method which produces extracellular deposits in the inner ear, whereas in the optic tectum extracellular precipitates are always present except with the oxalate-pyroantimonate procedure. Two factors have an influence on the reaction product: the duration of fixation and the type of resin. The prolongation of the fixation time up to 24 hours leads to an increase of the reaction product, which also becomes coarse-grained. These observations are corroborated by quantification with image analysis. Furthermore the use of an epoxy resin compared to acrylic resins decreases the amount of reaction product produced. We show that the application of several methods is meaningful in order to understand the calcium properties of the investigated tissue, but it is necessary to optimize a certain method for a given tissue.

Neuron-specific enolase as a marker of hypothalamo-neurohypophyseal development in postnatal Monodelphis domestica (Marsupialia)

In the newborn Monodelphis domestica, pituitary and brain are far from their definite shape and structural organization. The presumptive neurohypophysis appears as a mass of proliferating cells immunonegative for neuron-specific enolase (NSE) in the newborn, while perikarya positive for NSE are present in the rostroventral area of the hypothalamus. At the same time, the periventricular areas of the brain, consisting of proliferating cells, are non-reactive for NSE. In the 2.5-day-old specimens of this study, the neurohypophysis displays anti-NSE reactions in the peripheral regions. In animals 8–10 days of age, the amount of reaction product in the neurohypophysis has reached highest levels. This dramatic increase parallels the increase of NSE expression in perikarya of the rostroventral and caudoventral hypothalamus as well as the formation of a band-like reaction zone between the respective hypothalamic areas and the neurohypophysis.

Significance of high glucose-6-phosphatase activity in rat oviduct epithelium.

To study the origin of glucose in the oviduct fluid, we cytochemically examined glucose-6-phosphatase (G6Pase) activity in rat oviduct. The activity in the whole oviduct was also assayed biochemically. During proestrous, estrous, and metestrous phases, staining reaction for the activity was moderate in the epithelium of the caudal isthmus (CaI) and uterotubal junction (UJ), whereas it was weak in that of the ampulla (A) and cephalic isthmus (CeI). In the diestrous phase, staining reaction in the epithelium of CaI and UJ became strong although it remained weak in that of A and CeI. Reaction product for the activity was localized in the endoplasmic reticulum and nuclear envelope of all cell types in the epithelium. The amount of reaction product in secretory cells was small to moderate in CaI and UJ, and small in A and CeI during proestrus, estrus, and metestrus. In diestrous the amount became abundant in CaI and UJ and moderate in A and CeI. However, the amount in ciliated cells remained small in the four segments during the four phases. The biochemical activity in diestrous was greater than that in proestrus, estrus, or metestrus. This shows that the activity is high in secretory cells in the epithelium of CaI and UJ in the diestrous phase and suggests that the role of the high activity is to release glucose into the oviduct fluid for use by the embryo passing down the CaI and UJ to the uterus.

A quantitative enzyme histochemical analysis of the distribution of alkaline phosphatase activity in the periodontal ligament of the rat incisor.

The spatial distribution of alkaline phosphatase (ALP) activity was examined in the periodontal ligament of the continuously growing rat incisor. With the indoxyl-tetrazolium salt method, enzyme activity was demonstrated in undecalcified cryosections, and the amount of reaction product was quantified. ALP activity appeared to be distributed heterogeneously. Its highest activity was found in the bone-related compartment of the ligament. In the tooth-related compartment and the supracrestal extension of the ligament, enzyme activity was significantly lower, but still higher than in the lamina propria of the gingiva. In the part of the ligament bordering the cementum, highest activity was found in the apical region just occlusal to Hertwig's epithelial root sheath, where formation of acellular cementum begins. From there toward the incisal edge, the activity of the enzyme gradually decreased. It is suggested that differences among the various parts of the periodontal ligament are related to local variations in phosphate metabolism and cementum deposition.