Study of osteogenesis in adult sheep on a high and a low calcium ration by means of fluorescing bone markers

The rate of osteogenesis was studied in 8 non-pregnant, non-lactating Friesian dairy sheep, 3–6 years old, by means of a treatment with 3 different bone seeking agents. Four sheep were fed a low calcium ration (LCa : 1.8 g Ca/d) and four other sheep a high calcium ration (HCa : 12.7 g Ca/d). The bone markers, oxytetracycline-HCl, alizarine-complexon and demeclocycline-HCl, were administred at intervals of 6 weeks, and the sheep were killed 1 week after administration of the last marker. In undecalcified cross sections from the middle of ribs 2, 10 and 12, and from the proximal and distal parts of rib 10, the numbers of labelled osteons and the number of osteons with 1, 2 or 3 markers were counted under fluorescent microscopy.In the ribs of sheep from the LCa group, the number of labelled osteons and the quantity of labels per osteon tended to be higher than those of sheep from the HCa group. When osteogenic activity was compared in the different sites of ribs analysed, lowest osteogenic activity was observed in the proximal part of the 10th rib.The use of fluorescing markers offers the possibility of studying osteogenic activity over a certain period of time in adult sheep.     

Identification of several stilbene derivatives in bovine urine by means of high performance liquid chromatographic fractionation and immunochemical detection

Radioimmunochemical detection (RIA) following fractionation of urine extracts via high performance liquid chromatography (HPLC) turned out to be a very specific method for the identification of stilbene derivatives in bovine urine. Combination of the high resolution of the HPLC with a specific RIA is a suitable method to discriminate between the presence of different stilbene derivatives like diethylstilboestrol (DES), dienoestrol (DE) and hexoestrol (HEX) or other unknown compounds interfering in the celite-RIA used in the screening. Using this screening method 8200 samples of bovine urine were investigated on the presence of stilbene derivatives of which 133 were classified as 'positive'. In 106 'positive' urines the presence of DES was shown and in 19 'positive' urines the presence of DE or HEX, using the method described in this report whereas in 8 'positive' urines an unknown immunochemical active compound was detected. During 1.5 year of comparative investigation no qualitative discrepancies occurred between the results of the HPLC-immunogram procedure and the final confirmation by high resolution gas chromatography-mass spectometry (GCMS).     

Comparison of laboratory techniques for the evaluation of the antigenic potency of foot-and-mouth disease virus cultures and vaccines

Forty-seven suspensions of type A virus grown in surviving bovine tongue epithelium (Frenkel) cultures were compared in a quantitative complement fixation test (CFT) and an antibody combining test (ACT) to evaluate the antigenic mass of the 140 S component, and with three assay methods for infectivity titration.The cultures were then converted into vaccines and the potency of these was measured with a guinea-pig PD₅₀ test and again with the ACT. The relationship between the results obtained with different methods used for titration of infectivity, i.e. baby mice and plaque counting in BHK cells in monolayer (BHK-M) or suspended in agar (BHK-S) was poor and, with the exception of the BHK-S technique, showed little correlation with the results of the serological tests.The correlation coefficient between values obtained by the CFT and the ACT on virus cultures was 0.82 and that between CFT and ACT on vaccines 0.88. The guinea-pig PD₅₀'s of 19 vaccines were compared with the ACT and with the CFT and ACT of the corresponding cultures yielding r values of 0.74, 0.62 and 0.62 respectively. Regression lines are presented for the different relations.An accurate cattle PD₅₀ was determined for two vaccines of the series, using groups of 20 animals per vaccine dilution. The relationship between guinea-pig PD₅₀ (X) and cattle PD₅₀ (Y), expressed by the regression formula Y = 1.05 X + 0.84, showed that differences in cattle PD₅₀'s are of the same magnitude as those observed in the guinea-pig test, provided that sufficient numbers of animals are used for both tests.The results of the quantitative CFT and ACT were in good agreement with the guinea-pig test and as such these tests can provide valuable information on the quality of Frenkel cultures.     

Shifts in disease resistance induced by growth regulators

Experiments on the effect of various compounds with plant growth regulating activity led to the hypothesis that conditions inhibitory to indoleacetic acid (IAA) action or leading to a decrease in the IAA level in cucumber seedlings would be unfavourable for the development of cucumber scab, caused byCladosporium cucumerinum. Susceptibility decreased as the result of treatment with growth retardants, which cause an increase in IAA-oxidase activity in the plants, whereas application of indoleacetic acid or compounds expected to decrease the rate of oxidative breakdown of IAA in the seedlings resulted in an increase of susceptibility. Growing the plants under various periods of illumination also influenced both the susceptibility of the plants and the IAA-oxidase activity in the hypocotyl tissue.To obtain further information about the relation between the increase in resistance and the increase in the rate of IAA oxidation, the effect of one compound, viz. L-threo-β-phenylserine (abbr. phenylserine) was studied in more detail. The rate of oxidative breakdown of IAA was increased in extracts of hypocotyls and cotyledons of plants treated with phenylserine as compared with extracts of tissue of untreated plants. Extracts of phenylserine-treated plants were found to contain a higher cofactor and/or a lower inhibitor content than those of control plants. The substances involved were not identified, but this result may indicate a shift in the concentrations of various phenols in the tissue.The question must remain open whether a shift in the concentration of phenols some-how effected by phenylserine treatment determines both degree of susceptibility and rate of IAA oxidation independently, or whether the rate of IAA breakdown and the resulting change in IAA content in the plants is directly related to the degree of susceptibility.     

The energy metabolism of the adult common liver fluke,Fasciola hepatica

In adultFasciola hepatica glucose is broken down to volatile fatty acids, carbon dioxide and lactate. In the cytosol glucose is converted into malate and lactate and malate is metabolized further in the mitochondrial compartment via a dismutation. The dismutation of malate can proceed under anaerobic conditions. To a limited extent oxygen can function as terminal electron acceptor.It is shown that the reaction catalysed by fumarate reductase is a reversible reaction. The direction of the reaction depends on the relative concentrations of fumarate and succinate.Mechanisms of ATP production and electron transport under aerobic and anaerobic conditions are discussed. It is concluded that the energy metabolism of the adult liver fluke is adapted to the life in an environment with a low oxygen content.

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Kurzfassung Beim adultenFasciola hepatica wird Glucose in flüchtige Fettsäuren, Kohlendioxid und Laktate gespalten. Im Zytosol wird die Glucose in Malate und Laktate gespalten und Malate werden im Mitochondrienbereich durch Redox-Dismutation weiter verstoffwechselt. Die Dismutation der Malate kann unter anaeroben Bedingungen ablaufen. In gewissem Ausmass kann Sauerstoff terminal Elektronen binden.Es wird gezeigt, dass die Reaktion, die durch Fumaratreduktase katalysiert wird, reversibel ist. Der Ablauf der Reaktion hängt von den relativen Konzentrationen der Fumarate und Succinate ab.Ferner werden die Mechanismen der ATP-Produktion und des Elektronentransports unter aeroben und anaeroben Bedingungen diskutiert. Zusammenfassend wird festgestellt, dass der Energiestoffwechsel des adulten grossen Leberegels an ein Leben in einer sauerstoffarmen Umwelt adaptiert ist.

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Resume Chez les douves adultes le glucose est dégradé en acides gras volatil, gaz carbonique et lactate. Dans le cytosol le glucose est convertit en malate et en lactate et le malate est métabolisé ensuite dans la mitochondrie selon un phénomène de transformation biochimique. La transformation biochimique du malate peut se réaliser dans des conditions anaérobies. Dans une certaine mesure l'oxygène peut fonctionner comme un accepteur terminal d'électrons.Il est démontré que la réaction catalysée par la fumarate réductase est réversible. Le sens de la réaction dépend de la concentration relative en fumarate et succinate.Le mécanisme de production de l'ATP, du transport des électrons dans des conditions aérobies et anaérobies est discuté. Il est retenu que le métabolisme énergétique de la douve adulte est adapté à la vie dans un environnement pauvre en oxygène.

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Riassunto Negli adulti diFasciola hepatica il glucosio si scinde in acidi grassi volatili, biossido di carbonio e lattato. Nel citosol il glucosio è convertito in malato e lattato, ed il malato è ulteriormente metabolizzato nel comparto mitocondriale attraverso un processo di dismutazione. La dismutazione del malato può proseguire in condizioni di anaerobiosi. In misura limitata l'ossigeno può agire come accettore terminale di elettroni.E'stato dimostrato che la reazione catalizzata dalla fumarato-reduttasi è una reazione reversibile. La direzione della reazione dipende dalle concentrazioni relative di fumarato e di succinato.Vengono discussi i meccanismi di produzione di ATP e del trasporto di elettroni in condizione di aerobiosi e di anaerobiosi. Si conclude che il metabolismo energetico dell'adulto diFasciola hepatica è adattato alla vita in un ambiente con bassi livelli di ossigenazione.