53 research outputs found
Absence of Metabolic Cross-correction in Tay-Sachs Cells: IMPLICATIONS FOR GENE THERAPY
We have investigated the ability of a receptor-mediated gene transfer strategy (cross-correction) to restore ganglioside metabolism in fibroblasts from Tay-Sachs (TS) patients in vitro. TS disease is a GM2 gangliosidosis attributed to the deficiency of the lysosomal enzyme beta-hexosaminidase A (HexA) (beta-N-acetylhexosaminidase, EC ). The hypothesis is that transduced cells overexpressing and secreting large amounts of the enzyme would lead to a measurable activity in defective cells via a secretion-recapture mechanism. We transduced NIH3T3 murine fibroblasts with the LalphaHexTN retroviral vector carrying the cDNA encoding for the human Hex alpha-subunit. The Hex activity in the medium from transduced cells was approximately 10-fold higher (up to 75 milliunits) than observed in non-transduced cells. TS cells were cultured for 72 h in the presence of the cell medium derived from the transduced NIH3T3 cells, and they were analyzed for the presence and catalytic activity of the enzyme. Although TS cells were able to efficiently uptake a large amount of the soluble enzyme, the enzyme failed to reach the lysosomes in a sufficient quantity to hydrolyze the GM2 ganglioside to GM3 ganglioside. Thus, our results showed that delivery of the therapeutic HexA was not sufficient to correct the phenotype of TS cells
Anastomosis configuration and technique following ileocaecal resection for Crohn's disease: a multicentre study
A limited ileocaecal resection is the most frequently performed procedure for ileocaecal CD and different anastomotic configurations and techniques have been described. This manuscript audited the different anastomotic techniques used in a national study and evaluated their influence on postoperative outcomes following ileocaecal resection for primary CD. This is a retrospective, multicentre, observational study promoted by the Italian Society of Colorectal Surgery (SICCR), including all adults undergoing elective ileocaecal resection for primary CD from June 2018 May 2019. Postoperative morbidity within 30 days of surgery was the primary endpoint. Postoperative length of hospital stay (LOS) and anastomotic leak rate were the secondary outcomes. 427 patients were included. The side to side anastomosis was the chosen configuration in 380 patients (89%). The stapled anastomotic (n = 286; 67%), techniques were preferred to hand-sewn (n = 141; 33%). Postoperative morbidity was 20.3% and anastomotic leak 3.7%. Anastomotic leak was independent of the type of anastomosis performed, while was associated with an ASA grade ≥ 3, presence of perianal disease and ileocolonic localization of disease. Four predictors of LOS were identified after multivariate analysis. The laparoscopic approach was the only associated with a reduced LOS (p = 0.017), while age, ASA grade ≥ 3 or administration of preoperative TPN were associated with increased LOS. The side to side was the most commonly used anastomotic configuration for ileocolic reconstruction following primary CD resection. There was no difference in postoperative morbidity according to anastomotic technique and configuration. Anastomotic leak was associated with ASA grade ≥ 3, a penetrating phenotype of disease and ileo-colonic distribution of CD
National variations in perioperative assessment and surgical management of Crohn's disease: a multicentre study
Aim: Crohn's disease (CD) requires a multidisciplinary approach and surgery should be undertaken by dedicated colorectal surgeons with audited outcomes. We present a national, multicentre study, with the aim to collect benchmark data on key performance indicators in CD surgery, to highlight areas where standards of CD surgery excel and to facilitate targeted quality improvement where indicated. Methods: All patients undergoing ileocaecal or redo ileocolic resection in the participating centres for primary and recurrent CD from June 2018 to May 2019 were included. The main objective was to collect national data on hospital volume and practice variations. Postoperative morbidity was the primary outcome. Laparoscopic surgery and stoma rate were the secondary outcomes. Results: In all, 715 patients were included: 457 primary CD and 258 recurrent CD with a postoperative morbidity of 21.6% and 34.7%, respectively. Laparoscopy was used in 83.8% of primary CD compared to 31% of recurrent CD. Twenty-five hospitals participated and the total number of patients per hospital ranged from 2 to 169. Hospitals performing more than 10 primary CD procedures per year showed a higher adoption of laparoscopy and bowel sparing surgery. Conclusions: There is significant heterogeneity in the number of CD surgeries performed per year nationally in Italy. Our data suggest that high-volume hospitals perform more complex procedures, with a higher adoption of bowel sparing surgery. The rate of laparoscopy in high-volume hospitals is higher for primary CD but not for recurrent CD compared with low-volume hospitals
Semen molecular and cellular features: these parameters can reliably predict subsequent ART outcome in a goat model
Currently, the assessment of sperm function in a raw or processed semen sample is not able to reliably predict sperm ability to withstand freezing and thawing procedures and in vivo fertility and/or assisted reproductive biotechnologies (ART) outcome. The aim of the present study was to investigate which parameters among a battery of analyses could predict subsequent spermatozoa in vitro fertilization ability and hence blastocyst output in a goat model. Ejaculates were obtained by artificial vagina from 3 adult goats (Capra hircus) aged 2 years (A, B and C). In order to assess the predictive value of viability, computer assisted sperm analyzer (CASA) motility parameters and ATP intracellular concentration before and after thawing and of DNA integrity after thawing on subsequent embryo output after an in vitro fertility test, a logistic regression analysis was used. Individual differences in semen parameters were evident for semen viability after thawing and DNA integrity. Results of IVF test showed that spermatozoa collected from A and B lead to higher cleavage rates (0 < 0.01) and blastocysts output (p < 0.05) compared with C. Logistic regression analysis model explained a deviance of 72% (p < 0.0001), directly related with the mean percentage of rapid spermatozoa in fresh semen (p < 0.01), semen viability after thawing (p < 0.01), and with two of the three comet parameters considered, i.e tail DNA percentage and comet length (p < 0.0001). DNA integrity alone had a high predictive value on IVF outcome with frozen/thawed semen (deviance explained: 57%). The model proposed here represents one of the many possible ways to explain differences found in embryo output following IVF with different semen donors and may represent a useful tool to select the most suitable donors for semen cryopreservation
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A structural model for the assembly of the reaction centre and the B808-866 complex in the membranes of <i>Chloroflexus aurantiacus</i> based on the calculation of the triplet minus singlet spectrum of the primary donor
A well resolved triplet minus singlet spectrum of the primary donor P870 of the green bacterium Chloroflexus au- rantiacus, detected by optically detected magnetic resonance (ODMR), in intact membranes, was reported in our previous work [Photosynth. Res. 71 (2002) 45]. The spectrum is now interpreted and reconstructed by calculating the electronic interactions between the primary donor and the BChl a molecules belonging to the 866 absorption com- ponent of the B808-866 light-harvesting complex. The good agreement between the calculation and the experimental data strongly supports a topological model for the B808-866 complex which consists in a circular aggregate structure of a number of polypeptide units binding BChl a molecules assembled as in the LH2 peripheral antenna complex of purple bacteria, giving rise to two spectral forms. The aggregate surrounds the reaction centre (RC) resembling in this respect the LH1 core antenna complex of purple bacteria.</p
A structural model for the assembly of the reaction center and the B808-866 complex in the membranes of Chloroflexus aurantiacus based on the calculation of the Triplet-minus-Singlet spectrum of the primary donor
A well resolved triplet minus singlet spectrum of the primary donor P870 of the green bacterium Chloroflexus aurantiacus, detected by optically detected magnetic resonance (ODMR), in intact membranes, was reported in our previous work [Photosynth. Res. 71 (2002) 45]. The spectrum is now interpreted and reconstructed by calculating the electronic interactions between the primary donor and the BChl a molecules belonging to the 866 absorption component of the B808-866 light-harvesting complex. The good agreement between the calculation and the experimental data strongly supports a topological model for the B808-866 complex which consists in a circular aggregate structure of a number of polypeptide units binding BChl a molecules assembled as in the LH2 peripheral antenna complex of purple bacteria, giving rise to two spectral forms. The aggregate surrounds the reaction centre (RC) resembling in this respect the LH1 core antenna complex of purple bacteria. (C) 2003 Elsevier B.V. All rights reserved
Experimental evaluation of fatigue damage in two-stage loading tests based on the energy dissipation
Heat energy dissipation is a manifestation of damage accumulation in fatigue loaded components. Once recognized that some mechanical energy has to be expended to fatigue a material, energy partition into heat and stored energy is thought of as a material property. Since most of the mechanical input energy is dissipated into heat, the stored energy is difficult to be measured as difference between the expended and the dissipated energy. In the present paper heat energy is assumed as an index of fatigue damage. Since it reflects the material response to external loading, it was seen to reduce the scatter of constant amplitude fatigue test results with respect to the use of the stress amplitude. Moreover, two level fatigue test results could be interpreted with an higher level of accuracy when the Miner\u2019s rule was applied in terms of energy rather than stres
Fluorescence and absorption detected magnetic resonance of chlorosomes from green bacteria Chlorobium tepidum and Chloroflexus aurantiacus. A comparative study
A comparative study on the isolated chlorosomes from Chloriflexus aurantiacus, a green filamentous photosynthetic bacterium and Chlorobium tepidum, a green sulfur photosynthetic bacterium, was done by ODMR (optically detected magnetic resonance). Correlation between the results obtained by fluorescence and absorption detection is shown to be a sourer of information about the functional interactions among pigments. Analogies and differences are pointed out between the light-harvesting systems of the two species. Triplet states are easily detected in both bacteria at 1.8 K under steady-state illumination and are assigned to BChl c, BChl a, and carotenoid molecules. Carotenoids are found to be able to quench BChl a triplet states, but no evidence of BChl c triplet states quenching by this triplet-triplet transfer mechanism is found in both systems. Then from the data it appears that some carotenoids are in close contact with BChl a molecules. The relevance of this finding to the localization of carotenoids in the chlorosomes is discussed. in Cb, tepidum three different pools of BChl c oligomers connected to BChl a were found by detection of their triplet state, while only one pool of BChl c was evidenced in Cf. aurantiacus. The latter appears to be unconnected, at least at 1.8 K, to BChl a. On the other hand, heterogeneity in the BChl a triplet population was detected in Cf: aurantiacus. Even though the two bacteria show common features in the way the light excitation induces triplet formation at low temperature, the detected triplet states show spectroscopic properties that strongly depend on the system. The results clearly indicate that differences in pigment organization exist both in the fore and in the baseplate of the chlorosomes from the two different bacteria
Fluorescence and Absorption Detected Magnetic Resonance of Chlorosomes from Green Bacteria <i>Chlorobium tepidum</i> and <i>Chloroflexus aurantiacus</i>. A Comparative Study
A comparative study on the isolated chlorosomes from Chloroflexus aurantiacus, a green filamentous photosynthetic bacterium and Chlorobium tepidum, a green sulfur photosynthetic bacterium, was done by ODMR (optically detected magnetic resonance). Correlation between the results obtained by fluorescence and absorption detection is shown to be a source of information about the functional interactions among pigments. Analogies and differences are pointed out between the light-harvesting systems of the two species. Triplet states are easily detected in both bacteria at 1.8 K under steady-state illumination and are assigned to BChl c, BChl a, and carotenoid molecules. Carotenoids are found to be able to quench BChl a triplet states, but no evidence of BChl c triplet states quenching by this triplet−triplet transfer mechanism is found in both systems. Then from the data it appears that some carotenoids are in close contact with BChl a molecules. The relevance of this finding to the localization of carotenoids in the chlorosomes is discussed. In Cb. tepidum three different pools of BChl c oligomers connected to BChl a were found by detection of their triplet state, while only one pool of BChl c was evidenced in Cf. aurantiacus. The latter appears to be unconnected, at least at 1.8 K, to BChl a. On the other hand, heterogeneity in the BChl a triplet population was detected in Cf. aurantiacus. Even though the two bacteria show common features in the way the light excitation induces triplet formation at low temperature, the detected triplet states show spectroscopic properties that strongly depend on the system. The results clearly indicate that differences in pigment organization exist both in the core and in the baseplate of the chlorosomes from the two different bacteria.</p
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