Détermination quantitative des divers composants protéiques du suc pancréatique de porc. Etude du « stress » post opératoire, des variations nychtémérales et de l'adaptation à divers régimes alimentaires

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Starch digestion in tropical fishes : isolation, structural studies and inhibition kinetics of alpha-amylases from two tilapias Oreochromis niloticus and Sarotherodon melanotheron

Alpha-amylases from the intestinal cavity of two tilapia species, #Oreochromis niloticus (ONI-AMY) and #Sarotherodon melanotheron (SME-AMY), were purified using ammonium sulfate precipitation, affinity chromatography and chromatofocusing procedures. The purification was approximately 100-fold. The amylolytic activity, specific activity, product distribution, pH and temperature profile of ONI-AMY and SME-AMY are quite similar. The molecular mass differs slightly : 56 600 (ONI-AMY) vs. 55 500 (SME-AMY). As shown by isoelectric focusing analysis, both amylases contain two isoforms A and B with distinct pI : 7.2 (A) and 7.8 (B), vs. 8.3 (A) and 8.8 (B), respectively. It was not possible to isolate B, since B converts into A with time. The kinetics of the inhibition of ONI-AMY and SME-AMY activity by alpha, Beta, and gamma-cyclodextrin (alpha-, Beta- and gamma-CD) were investigated using amylose as the substrate. Statistical analysis of the kinetic data expressed using a general velocity equation and assuming rapid equilibrium showed that the inhibition is of the mixed noncompetitive type. Similar results were obtained with ONI-AMY and SME-AMY. Beta- and gamma-CD are stronger inhibitors than alpha-CD. ONI-AMY and SME-AMY are then closely related and show the general features common to the members of the alpha-amylase class (family 13). They enable ONI and SME tilapias to digest starch in food. (Résumé d'auteur

Fistula plug in fistulising ano-perineal Crohn's disease: a randomised controlled trial

International audienceBackground and aims: Anal fistula plug (AFP) is a bioabsorbable bioprosthesis used in ano-perineal fistula treatment. We aimed to assess efficacy and safety of AFP in fistulising ano-perineal Crohn's disease (FAP-CD). Methods: In a multicentre, open-label, randomized controlled trial we compared seton removal alone (control group) with AFP insertion (AFP group) in 106 Crohn's disease patients with non or mildly active disease having at least one ano-perineal fistula tract drained for more than one month. Patients with abscess (collection ≥ 3 mm on MRI) or recto-vaginal fistulas were excluded. Randomization was stratified in simple or complex fistulas according to AGA classification. Primary end point was fistula closure at week 12.Results54 patients were randomized to AFP group (control group 52). Median fistula duration was 23 [10-53] months. Median Crohn's Disease Activity Index at baseline was 81 [45-135]. Fistula closure at week 12 was achieved in 31.5% patients in AFP group and in 23.1 % in control group (relative risk stratified on AGA classification, RR: 1.31; 95%CI: 0.59-4.02; p=0.19). No interaction in treatment effect with complexity stratum was found. 33.3% patients with complex fistula and 30.8% patients with simple fistula closed the tracts after AFP, as compared to 15.4% and 25.6% in controls respectively (RR of success=2.17 in complex fistula vs. RR=1.20 in simple fistula; p= 0.45). Concerning safety, at week 12, 17 patients developed at least one adverse event in AFP group vs. 8 in controls (p=0.07). Conclusion: AFP is not more effective than seton removal alone to achieve FAP-CD closur

All-Inside Bicruciate Ligament Reconstruction Technique: A Focus on Graft Tensioning Sequence

Bicruciate ligament (BCL) reconstructions are challenging procedures. One of the main operative goals is to stabilize the knee in the correct anterior-posterior position. We present an all-inside arthroscopic BCL reconstruction technique using hamstring tendon grafts. Ipsilateral semitendinosus (ST) and gracilis tendons are used for TriLink (Arthrex, Naples, FL) double-bundle posterior cruciate ligament (PCL) reconstruction and contralateral ST tendon is used for GraftLink (Arthrex) single-bundle anterior cruciate ligament (ACL) reconstruction. The use of instruments for retrograde reaming and devices for adjustable cortical suspensory fixation allows for a safe, reproducible all-inside BCL reconstruction by simplifying these difficult steps. To minimize the risk of anterior-posterior malposition, the ACL graft is first tensioned with the knee in full extension, ensuring a neutral anteroposterior positioning of the tibia under the femur. The PCL anterolateral bundle can then be independently tensioned with the knee at 90° of flexion, and the posteromedial bundle at 30° of flexion, while applying an anterior translation to the tibia to reduce the posterior drawer without any risk of overcorrection. The purpose of this Technical Note was to describe an all-inside BCL reconstruction with a specific focus on the graft tensioning sequence

Gene transfer of integration defective anti-HSV-1 meganuclease to human corneas ex vivo

Corneal graft rejection is a major problem in chronic herpetic keratitis (HK) patients with latent infection. A new class of antiviral agents targeting latent and active forms of herpes simplex virus type 1 (HSV-1) is importantly required. Meganucleases are sequence-specific homing endonucleases capable of inducing DNA double-strand breaks. A proof-of-concept experiment has shown that tailor-made meganucleases are efficient against HSV-1 in vitro. To take this work a step forward, we hypothesized that the pre-treatment of human corneas in eye banks using meganuclease-encoding vectors will allow HK patients to receive a medicated cornea to resist the recurrence of the infection and the common graft rejection problem. However, this strategy requires efficient gene delivery to human corneal endothelium. Using recombinant adeno-associated virus, serotype 2/1 (rAAV2/1), efficient gene delivery of a reporter gene was demonstrated in human corneas ex vivo. The optimum viral dose was 3.7 7 10(11) VG with an exposure time of 1 day, followed by 6 days incubation in de-swelling medium. In addition, 12 days incubation can result in transgene expression in excess of 70%. Using similar transduction conditions, meganuclease transgene expression was detected in 39.4% of the endothelial cells after 2 weeks in culture. Reduction of the total viral load in the media and the endothelial cells of corneas infected with HSV-1 was shown. Collectively, this work provides information about the optimum conditions to deliver genetic material to the cornea, and demonstrates for the first time the expression of meganuclease in human corneas ex vivo and its antiviral activity. In conclusion, we demonstrate that the treatment of human corneas in eye banks before transplantation is a new approach to address the unmet clinical needs in corneal diseases

Measurement of the displacement field of dislocations to 0.03 Å by electron microscopy

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