Integrated approach on heat transfer and inactivation kinetics of microorganisms on the surface of foods during heat treatments: Software development

The objective of this work was to create a software application (Bugdeath 1.0) for the simulation of inactivation kinetics of microorganisms on the surface of foods, during dry and wet pasteurisation treatments. The program was developed under the Real Basic 5.2 application, and it is a user-friendly tool. It integrates heat transfer phenomena and microbial inactivation under constant and time-varying temperature conditions. On the basis of the selection of a heating regime of the medium, the program predicts the food surface temperature and the change in microbial load during the process. Input data and simulated values can be visualised in graphics or data tables. Printing, exporting and saving file options are also available. Bugdeath 1.0 includes also a useful database of foods (beef and potato) and related thermal properties, microorganisms (Salmonella and Listeria monocytogenes) and corresponding inactivation kinetic parameters. This software can be coupled to an apparatus developed under the scope of the European Project BUGDEATH (QLRT-2001-01415), which was conceived to provide repeatable surface temperature-time treatments on food samples. The program has also a great potential for research and industrial applications

The establishment of the first reported zebrafish model for thoracic aortic dissection and rupture

About 0,16 percent of the Western population suffers from thoracic aortic aneurysms and dissections (TAAD). Weakening of the vessel wall of the thoracic aorta increases the risk for aortic dissection and rupture, which associates with a high mortality rate. Current treatment options in TAAD are limited to a pharmacological reduction of hemodynamic stress and surgical repair at a critical diameter. Despite the availability of different mouse models for TAAD, the underlying molecular mechanisms remain elusive. We therefore developed a zebrafish model for aortic dissection/rupture. For this purpose, we targeted 2 genes involved in angiogenesis, SMAD3 and SMAD6. In humans, loss of function (LOF) of SMAD3 results in TAAD, arterial tortuosity and early onset osteoarthritis. SMAD6 LOF mutations increase the risk for a bicuspid aortic valve and TAAD. In zebrafish, both SMAD3 and SMAD6 have 2 paralogues. Using CRISPR/Cas9 gene editing technology, we developed a quadruple knockout (KO): smad3a-/-;smad3b-/-;smad6a-/-;smad6b-/-. At 5 days post fertilization, quadruple KO embryos showed asymmetrical branching of the aortic arches. Survival of adult quadruple KO zebrafish was severely decreased and all quadruple mutants died before the age of one year. A stress-inducing protocol caused sudden death in 60% of the mutant zebrafish. Histochemical investigation of consecutive sections of the ventral aorta in quadruple mutants stained for elastin showed medial elastolysis, intramural hematomas, aortic dissections and ruptures, which was further supported by 3D reconstructions. These observations indicate that we successfully developed the first ever reported zebrafish model for aortic dissection/rupture. This model will be highly valuable to better understand the pathogenic processes underlying TAAD and to evaluate potential therapeutic compounds

The establishment of the first reported zebrafish model for thoracic aortic dissection and rupture

Per year, 3-4 persons per 100,000 suffer from thoracic aortic dissection (TAD), causing significant morbidity and mortality. Dissections occur often at sites where neural crest and mesodermal derived cell populations interact. In TAD, an interplay between hemodynamic stress, tissue remodeling due to impaired extracellular matrix assembly, aberrant growth factor signaling and cell-matrix mechanosensing weakens the vessel wall. Despite the availability of different mouse models for TAD, the cascade of underlying mechanisms remains largely elusive. Consequently, current treatment options are limited to a pharmacological reduction of hemodynamic stress and surgical repair at a critical diameter. We therefore developed a zebrafish model for aortic dissection/rupture targeting two genes involved in angiogenesis, SMAD3 and SMAD6. In humans, loss of function (LOF) of SMAD3 results in thoracic aortic aneurysm and dissection (TAAD), arterial tortuosity and early onset osteoarthritis. SMAD6 LOF mutations increase the risk for a bicuspid aortic valve and TAAD. In zebrafish, both SMAD3 and SMAD6 have 2 paralogues. Using CRISPR/Cas9 gene editing technology, we developed a quadruple knockout (KO): smad3a-/-;smad3b-/-;smad6a-/-;smad6b-/-. At 5 days post fertilization, quadruple KO embryos showed asymmetrical branching of the aortic arches. Survival of adult quadruple KO zebrafish was severely decreased and all but one quadruple mutants died before the age of one year. A stress-inducing protocol caused sudden death in 60% of the mutant zebrafish. Histology of consecutive sections of the ventral aorta in quadruple mutants stained for elastin showed medial elastolysis, intramural hematomas, aortic dissections and ruptures, which was further supported by 3D reconstructions. RNA sequencing revealed upregulation of melanogenesis as well as mitfa, an important transcription factor in neural crest, relevant for the pathogenesis. In conclusion, we successfully developed the first zebrafish model for aortic dissection/rupture. This model will be highly relevant to better understand the pathogenesis underlying TAD and to evaluate potential therapeutic compounds

The establishment of the first reported zebrafish model for thoracic aortic dissection and rupture

3-4 cases per 100,000 persons per year are estimated to suffer from thoracic aortic dissection (TAD). Weakening of the vessel wall of the thoracic aorta increases the risk for TAD and rupture, which associates with a high mortality rate. Current treatment options in thoracic aortic dissections (TAAD) are limited to a pharmacological reduction of hemodynamic stress and surgical repair at a critical diameter. Despite the availability of different mouse models for TAD, the underlying molecular mechanisms remain elusive. We therefore developed a zebrafish model for aortic dissection/rupture. For this purpose, we targeted 2 genes involved in angiogenesis, SMAD3 and SMAD6. In humans, loss of function (LOF) of SMAD3 results in thoracic aortic aneurysm and dissection (TAAD), arterial tortuosity and early onset osteoarthritis. SMAD6 LOF mutations increase the risk for a bicuspid aortic valve and TAAD. In zebrafish, both SMAD3 and SMAD6 have 2 paralogues. Using CRISPR/Cas9 gene editing technology, we developed a quadruple knockout (KO): smad3a-/-;smad3b-/-;smad6a-/-;smad6b-/-. At 5 days post fertilization, quadruple KO embryos showed asymmetrical branching of the aortic arches. Survival of adult quadruple KO zebrafish was severely decreased and all but one quadruple mutants died before the age of one year. A stress-inducing protocol caused sudden death in 60% of the mutant zebrafish. Histochemical investigation of consecutive sections of the ventral aorta in quadruple mutants stained for elastin showed medial elastolysis, intramural hematomas, aortic dissections and ruptures, which was further supported by 3D reconstructions. RNA sequencing revealed upregulation of melanogenesis as well as upregulation of transcription factor mitfa, which might be involved in the pathogenesis. These observations indicate that we successfully developed the first ever reported zebrafish model for aortic dissection/rupture. This model will be highly valuable to better understand the pathogenic processes underlying TAAD and to evaluate potential therapeutic compounds

The establishment of the first reported zebrafish model for thoracic aortic dissection and rupture

3-4 cases per 100,000 persons per year are estimated to suffer from thoracic aortic dissection (TAD). Weakening of the vessel wall of the thoracic aorta increases the risk for TAD and rupture, which associates with a high mortality rate. Current treatment options are limited to a pharmacological reduction of hemodynamic stress and surgical repair at a critical diameter. Despite the availability of different mouse models for TAD, the underlying molecular mechanisms remain elusive. We therefore developed a zebrafish model for aortic dissection/rupture. For this purpose, we targeted 2 genes involved in angiogenesis, SMAD3 and SMAD6. In humans, loss of function (LOF) of SMAD3 results in thoracic aortic aneurysm and dissection (TAAD), arterial tortuosity and early onset osteoarthritis. SMAD6 LOF mutations increase the risk for a bicuspid aortic valve and TAAD. In zebrafish, both SMAD3 and SMAD6 have 2 paralogues. Using CRISPR/Cas9 gene editing technology, we developed a quadruple knockout (KO): smad3a-/-;smad3b-/-;smad6a-/-;smad6b-/-. Survival of adult quadruple KO zebrafish was severely decreased. All but one quadruple mutant died before the age of one year. A stress-inducing protocol caused sudden death in 60% of the mutant zebrafish. Histochemical investigation of consecutive sections of the ventral aorta in quadruple mutants stained for elastin showed medial elastolysis, intramural hematomas, aortic dissections and ruptures, which was further supported by 3D reconstructions. RNA sequencing revealed upregulation of melanogenesis and transcription factor mitfa, which might be involved in the pathogenesis. These observations indicate that we successfully developed the first ever reported zebrafish model for aortic dissection/rupture

The establishment of the first reported zebrafish model for thoracic aortic dissection and rupture

With 3-4 cases per 100,000 person-years, thoracic aortic dissection (TAD) is a relatively rare but devastating disease that associates with a high mortality rate. Weakening of the vessel wall and progressive dilatation of the thoracic aorta may precede TAD, but often remains undetected. Treatment options are limited and consist of surgical repair at the critical diameter as there is currently no pharmacological intervention available. Despite the existence of different mouse models for TAD, the underlying disease mechanisms remain largely elusive. We developed a zebrafish model for aortic dissection/rupture targeting 2 genes involved in angiogenesis, SMAD3 and SMAD6. In humans, loss of function (LOF) of SMAD3 results in thoracic aortic aneurysm and dissection (TAAD), arterial tortuosity and early onset osteoarthritis. SMAD6 LOF mutations increase the risk for a bicuspid aortic valve and TAAD. In zebrafish, both SMAD3 and SMAD6 have 2 paralogues. Using CRISPR/Cas9 gene-editing technology, we developed a quadruple knockout (KO): smad3a-/-;smad3b-/-;smad6a-/-;smad6b-/-. Survival of adult quadruple KO zebrafish is severely decreased (<1 year). A stress-inducing protocol causes sudden death in 60% of the mutant zebrafish. Histochemical investigation of consecutive sections of the ventral aorta in quadruple mutants stained for elastin shows medial elastolysis, intramural hematomas, false lumens and aortic dissections and ruptures at sites with high hemodynamic stress, as supported by 3D-reconstructions. RNA-sequencing reveals upregulation of melanogenesis and the transcription factor mitfa, a previously unaddressed athway. Hence, we successfully developed the first-ever reported zebrafish model for TAD that reveals unexpected novel mechanistic insights in TAD, targetable for therapy