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The inhibition effect of cellobiose on the initial stage of hydrolysis when cellobiohydrolase Cel 7A and endoglucanases Cel 7B, Cel 5A, and Cel 12A from Trichoderma reesei were acting on bacterial cellulose and amorphous cellulose that were [(3)H]- labeled at the reducing end was quantified. The apparent competitive inhibition constant (K(i)) for Cel 7A on [(3)H]-bacterial cellulose was found to be 1.6 +/- 0.5 mM, 100-fold higher than that for Cel 7A acting on low-molecular-weight model substrates. The hydrolysis of [(3)H]-amorphous cellulose by endoglucanases was even less affected by cellobiose inhibition with apparent K(i) values of 11 +/- 3 mM and 34 +/- 6 mM for Cel 7B and Cel 5A, respectively. Contrary to the case for the other enzymes studied, the release of radioactive label by Cel 12A was stimulated by cellobiose, possibly due to a more pronounced transglycosylating activity. Theoretical analysis of the inhibition of Cel 7A by cellobiose predicted an inhibition analogous to that of mixed type with two limiting cases, competitive inhibition if the prevalent enzyme-substrate complex without inhibitor is productive and conventional mixed type when the prevalent enzyme-substrate complex is nonproductive.

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BackgroundSurgical castration in male piglets is painful and methods that reduce this pain are requested. This study evaluated the effect of local anaesthesia and analgesia on vocal, physiological and behavioural responses during and after castration. A second purpose was to evaluate if herdsmen can effectively administer anaesthesia.MethodsFour male piglets in each of 141 litters in five herds were randomly assigned to one of four treatments: castration without local anaesthesia or analgesia (C, controls), analgesia (M, meloxicam), local anaesthesia (L, lidocaine), or both local anaesthesia and analgesia (LM). Lidocaine (L, LM) was injected at least three minutes before castration and meloxicam (M, LM) was injected after castration. During castration, vocalisation was measured and resistance movements judged. Behaviour observations were carried out on the castration day and the following day. The day after castration, castration wounds were ranked, ear and skin temperature was measured, and blood samples were collected for analysis of acute phase protein Serum Amyloid A concentration (SAA). Piglets were weighed on the castration day and at three weeks of age. Sickness treatments and mortality were recorded until three weeks of age.ResultsPiglets castrated with lidocaine produced calls with lower intensity (p < 0.001) and less resistance movements (p < 0.001) during castration. Piglets that were given meloxicam displayed less pain-related behaviour (huddled up, spasms, rump-scratching, stiffness and prostrated) on both the castration day (p = 0.06, n.s.) and the following day (p = 0.02). Controls had less swollen wounds compared to piglets assigned to treatments M, L and LM (p < 0.001). The proportion of piglets with high SAA concentration (over threshold values 200, 400 mg/l) was higher (p = 0.005; p = 0.05) for C + L compared to M + LM. Ear temperature was higher (p < 0.01) for controls compared to L and LM. There were no significant treatment effects for skin temperature, weight gain, sickness treatments or mortality.ConclusionsThe study concludes that lidocaine reduced pain during castration and that meloxicam reduced pain after castration. The study also concludes that the herdsmen were able to administer local anaesthesia effectively.

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