The enteric reflexes in isolated segments of the distal colon and rectum of the guinea-pig were studied by applying localized distensions and recording the consequent changes in circular muscle activity, and by recording tension changes in the circular muscle during the propulsion of a bolus in vitro. Lesions of the wall of the colon were made to locate nerve pathways involved in the reflexes and pharmacological tests were applied to investigate the natures of transmitters released and the types of receptors involved. Distension produced a transient contraction of the circular muscle on the oral side and sustained relaxation on the anal side. Both reflexes were nerve-mediated. They were elicited in segments deprived of mucosa and submucosa. Interruption of Auerbach's plexus, but not interruption of the submucosal plexus, prevented their conduction. The ascending excitatory reflex was partly blocked by hyoscine and was also partly blocked by methysergide or by making the preparation tachyphylactic to the excitatory action of 5-hydroxytryptamine. The ascending excitatory pathways apparently involve neurons releasing a 5-HT-like transmitter as well as cholinergic neurons. The descending inhibitory reflex was not antagonized by hyoscine, guanethidine, methysergide or mepyramine. It is assumed that the inhibitory neurons activated in this reflex are identical with the non-cholinergic, non-adrenergic, enteric inhibitory neurons found throughout the intestine. If both the ascending excitatory and descending inhibitory reflexes acted simultaneously on the same area of circular muscle, the inhibitory response tended to dominate. Pellets of faeces, covered by a thin layer of resin, were introduced into the oral ends of isolated segments of colon. They were propelled analwards at speeds of 0.5-1.6 mm/s. Tension records showed that the pellets were preceded by relaxation and followed by a ring of contraction. The propulsion was blocked by both hyoscine and methysergide. Descending waves of contraction were also observed in empty segments of colon. These occurred spontaneously or were initiated by stretch. They did not occur in the presence of hyoscine or tetrodotoxin. It is postulated that three factors may contribute to propulsion in the guinea-pig distal colon: ascending excitatory reflexes which evoke contractions above a bolus; descending inhibitory reflexes which cause relaxations below; and contractions which, once set up in the circular muscle, travel in an anal direction.
The properties of a new fluorescence histochemical method for arylethylamines based on reaction with a mixture of 4% formaldehyde and 0.5% glutaraldehyde in aqueous solution are described. At room temperature the aldehyde mixture produced a well-localized fluorescence reaction in tissues, which, when examined microscopically in aqueous solution, was sufficiently intense for fine terminal noradrenergic axons to be seen. If the tissue was subsequently dried, the fluorescence intensity increased. At the same time as inducing the fluorophores, the aldehyde mixture fixed the tissue to a standard well suited for electron microscopy. It thus proved possible to locate amine containing cells in the fluorescence microscope and subsequently examine their ultrastructure. In aqueous models, the aldehyde mixture formed fluorescent products with adrenaline, noradrenaline, dopamine, dopa, 5-hydroxytryptamine and 5-hydroxytryptophan, but not with histamine or octopamine. The fluorescence induced in the aldehyde mixture remained stable if the tissue was subsequently transferred to saline or distilled water and when it was dehydrated in ethanol and cleared with xylene, benzene, chloroform or acetone.
The reactions of glyoxylic acid with peripheral stores of noradrenaline and 5-hydroxytryptamine to provide a fluorescence histochemical method for their localization have been investigated. Incubation in glyoxylic acid, followed by drying and heating of whole mount preparations gives an intense and well localized reaction. For incubation, a concentration of 2% glyoxylic acid, buffered to pH 7 at room temperature for 30 minutes gives ideal results. The method is equally good if the pH is varied in the range 6 to 9 or if the tissue is stored in the incubation mixture for up to 6 hours. Ideal development of the fluorophore requires an initial excess of moisture in the tissue, that this moisture is driven off during development, and that the tissue is protected from further moistening. A suitable method of achieving these ends is to heat partially dried tissue at 100 degrees C for 4 minutes and then cover it with paraffin oil. 5-hydroxytryptamine can be readily distinguished from noradrenaline because it forms a fluorophore after reaction at pH 3.5, whereas noradrenaline does not. Both amines can be visualized after incubation at neutral pH. Comparison with the formaldehyde vapour technique reveals three main advantages (and no disadvantages) of the glyoxylic acid method: (1) it gives a finer localization with higher fluorescence yield, (2) the glyoxylic acid method is less susceptible to variations in procedure and, (3) it is both simpler and quicker to apply.
This study investigates the influence of chemical composition on the biochemical methane potential (BMP) of twelve different batches of fruit and vegetable waste (FVW) with different compositions collected over one year. BMP ranged from 288 to 516LCHkgVS, with significant statistical differences between means, which was explained by variations in the chemical composition over time. BMP was most strongly correlated to lipid content and high calorific values. Multiple linear regression was performed to develop statistical models to more rapidly predict methane potential. Models were analysed that considered chemical compounds and that considered only high calorific value as a single parameter. The best BMP prediction was obtained using the statistical model that included lipid, protein, cellulose, lignin, and high calorific value (HCV), with R of 92.5%; lignin was negatively correlated to methane production. Because HCV and lipids are strongly correlated, and because HCV can be determined more rapidly than overall chemical composition, HCV may be useful for predicting BMP.
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