Effect of glycerol, n, n-dimethylformamide and n-methyl-2-pyrrolidone on rabbit sperm stored at 4 °C and 16 °C

Artificial insemination with cooled semen is the most common practice in rabbit farms and any improvement on it helps to increase the efficiency and productivity of rabbit meat farms. Therefore, the aim of this study was to assess whether different cryoprotectant agents (CPA) as glycerol, N, N-Dimethylformamide (DMF) and N-Methyl--2-Pyrrolidone (NMP) can improve cooled rabbit sperm quality stored at 4C and 16C. Sperm samples were diluted with INRA 96® (Extender A), INRA 96® with 6% glycerol (Extender B) or 6% DMF (Extender C) or 6% NMP (Extender D) respectively and stored at 4C and 16C. Samples were then analysed at 4, 24, 48 and 72 hours after refrigeration by integrated sperm analysis system (ISAS), eosin-nigrosin stain (vitality), hypo-osmotic swelling test (HOS test) and acrosome integrity test. Extender C showed higher percentage of motility, vitality and HOS test than extender B and D (p<0.05). Whereas sperm quality decreased over time (p<0.05), data showed that the addition of DMF kept the motility and sperm plasma membrane integrity after 24 hours of storage better than other diluents. These results suggest that the addition of DMF to INRA 96® exerts a protective effect on the membrane of spermatozoa improving seminal quality

Long-term preservation of freeze-dried rabbit sperm by adding rosmarinic acid and different chelating agents

Freeze-drying (FD) technique has been applied as an alternative technology to preserve gene resources to allow simple sperm preservation and shipment at 4 degrees C. Nevertheless, DNA sperm might be damaged by mechanical or oxidative stress throughout FD procedure. Therefore, suitable protection to maintain DNA integrity is required. The aim of this study was to determine the effect of rosmarinic acid (RA) as an antioxidant and two chelating agents (EGTA and EDTA) on the DNA integrity of freeze-dried rabbit sperm after storage of the samples at 4 degrees C and room temperature for 8 months. Rabbit sperm were freeze-dried in basic medium (10 mM Tris-HCl buffer and 50 mM NaCl) supplemented with 50 mM EGTA (1), 50 mM EGTA plus 105 mu M RA (2), 50 mM EDTA (3) or 50 mM EDTA plus 105 mu M RA (4). Semen samples were kept at 4 degrees C and room temperature during 8 months. After rehydration, DNA integrity was evaluated with Sperm Chromatin Dispersion test observing that DNA fragmentation was higher when semen samples were freeze-dried with EGTA (10.9%) than with EDTA (4.1%) (p < 0.01). Furthermore, RA acted better under adverse conditions and no significant differences were found in temperature storage. Summarizing, FD is a method that can allow simple gene resources preservation among 4 degrees C to 25 degrees C during 8 months and transportation without the need for liquid nitrogen or dry ice. EDTA chelating agent is the most suitable media for freeze-dried rabbit sperm and the addition of RA protects the DNA against the oxidative stress caused during FD procedure

Struvite precipitation in wastewater treatment plants anaerobic digestion supernatants using a magnesium oxide by-product

Struvite precipitation is a well-known technology to recover and upcycle phosphorus from municipal wastewater as a slow-release fertiliser. However, the economic and environmental costs of struvite precipitation are constrained by using technical-grade reagents as a magnesium source. This research evaluates the feasibility of using a low-grade magnesium oxide (LG-MgO) by-product from the calcination of magnesite as a magnesium source to precipitate struvite from anaerobic digestion supernatants in wastewater treatment plants. Three distinct LG-MgOs were used in this research to capture the inherent variability of this by-product. The MgO content of the LG-MgOs varied from 42 % to 56 %, which governed the reactivity of the by-product. Experimental results showed that dosing LG-MgO at P:Mg molar ratio close to stoichiometry (i.e. 1:1 and 1:2) favoured struvite precipitation, whereas higher molar ratios (i.e. 1:4, 1:6 and 1:8) favoured calcium phosphate precipitation due to the higher calcium concentration and pH. At a P:Mg molar ratio of 1:1 and 1:2, the percentage of phosphate precipitated was 53-72 % and 89-97 %, respectively, depending on the LG-MgO reactivity. A final experiment was performed to examine the composition and morphology of the precipitate obtained under the most favourable conditions, which showed that (i) struvite was the mineral phase with the highest peaks intensity and (ii) struvite was present in two different shapes: hopper and polyhedral. Overall, this research has demonstrated that LG-MgO is an efficient source of magnesium for struvite precipitation, which fits the circular economy principles by valorising an industrial by-product, reducing the pressure on natural resources, and developing a more sustainable technology for phosphorus recovery

ALK2 inhibitors display beneficial effects in preclinical models of <i>ACVR1</i> mutant diffuse intrinsic pontine glioma.

Diffuse intrinsic pontine glioma (DIPG) is a lethal childhood brainstem tumour, with a quarter of patients harbouring somatic mutations in ACVR1, encoding the serine/threonine kinase ALK2. Despite being an amenable drug target, little has been done to-date to systematically evaluate the role of ACVR1 in DIPG, nor to screen currently available inhibitors in patient-derived tumour models. Here we show the dependence of DIPG cells on the mutant receptor, and the preclinical efficacy of two distinct chemotypes of ALK2 inhibitor in vitro and in vivo. We demonstrate the pyrazolo[1,5-a]pyrimidine LDN-193189 and the pyridine LDN-214117 to be orally bioavailable and well-tolerated, with good brain penetration. Treatment of immunodeprived mice bearing orthotopic xenografts of H3.3K27M, ACVR1R206H mutant HSJD-DIPG-007 cells with 25 mg/kg LDN-193189 or LDN-214117 for 28 days extended survival compared with vehicle controls. Development of ALK2 inhibitors with improved potency, selectivity and advantageous pharmacokinetic properties may play an important role in therapy for DIPG patients

Repurposing Vandetanib plus Everolimus for the Treatment of ACVR1-Mutant Diffuse Intrinsic Pontine Glioma.

Somatic mutations in ACVR1 are found in a quarter of children with diffuse intrinsic pontine glioma (DIPG), but there are no ACVR1 inhibitors licensed for the disease. Using an artificial intelligence-based platform to search for approved compounds for ACVR1-mutant DIPG, the combination of vandetanib and everolimus was identified as a possible therapeutic approach. Vandetanib, an inhibitor of VEGFR/RET/EGFR, was found to target ACVR1 (K d = 150 nmol/L) and reduce DIPG cell viability in vitro but has limited ability to cross the blood-brain barrier. In addition to mTOR, everolimus inhibited ABCG2 (BCRP) and ABCB1 (P-gp) transporters and was synergistic in DIPG cells when combined with vandetanib in vitro. This combination was well tolerated in vivo and significantly extended survival and reduced tumor burden in an orthotopic ACVR1-mutant patient-derived DIPG xenograft model. Four patients with ACVR1-mutant DIPG were treated with vandetanib plus an mTOR inhibitor, informing the dosing and toxicity profile of this combination for future clinical studies. SIGNIFICANCE: Twenty-five percent of patients with the incurable brainstem tumor DIPG harbor somatic activating mutations in ACVR1, but there are no approved drugs targeting the receptor. Using artificial intelligence, we identify and validate, both experimentally and clinically, the novel combination of vandetanib and everolimus in these children based on both signaling and pharmacokinetic synergies.This article is highlighted in the In This Issue feature, p. 275

Responsabilidad objetiva versus responsabilidad subjetiva : negación indebida de la defensa de falta de culpa

Introducción: La tesis que presento para el juicio de quienes tengan a su cargo su evaluación se basa en la consideración de un caso real –bien conocido por el autor– para, a partir de allí, llevar a cabo un estudio del alcance de la competencia sancionatoria de la Administración en el marco del ejercicio de su poder de policía. El trabajo se plantea a partir de los siguientes interrogantes: ¿Puede un funcionario responsable del Control y Prevención de Lavado de Dinero ser responsabilizado por el solo hecho de ocupar el cargo, sin imputación concreta de conducta irregular alguna? En su caso, ¿qué excusas absolutorias tiene para oponer a la pretensión punitiva? ¿Sería suficiente con probar que ha obrado con cuidado y previsión, dando órdenes precisas, capacitando al personal, monitoreando, poniendo los medios con debida diligencia, etc.? O, por el contrario, ¿puede ser penado aunque hubiera obrado bien? ¿Cuál es la responsabilidad de la persona jurídica? En un primer encuadre para responder a estas preguntas, partí de lo expresado por la Sala II de la Cámara Nacional de Apelaciones en lo Contencioso Administrativo Federal, que ha dicho que “la simple constatación de la comisión de infracciones genera la consiguiente responsabilidad y sanción del infractor, salvo que éste invoque y demuestre la existencia de una circunstancia exculpatoria válida”

Responsabilidad objetiva versus responsabilidad subjetiva : negación indebida de la defensa de falta de culpa

Introducción: La tesis que presento para el juicio de quienes tengan a su cargo su evaluación se basa en la consideración de un caso real –bien conocido por el autor– para, a partir de allí, llevar a cabo un estudio del alcance de la competencia sancionatoria de la Administración en el marco del ejercicio de su poder de policía. El trabajo se plantea a partir de los siguientes interrogantes: ¿Puede un funcionario responsable del Control y Prevención de Lavado de Dinero ser responsabilizado por el solo hecho de ocupar el cargo, sin imputación concreta de conducta irregular alguna? En su caso, ¿qué excusas absolutorias tiene para oponer a la pretensión punitiva? ¿Sería suficiente con probar que ha obrado con cuidado y previsión, dando órdenes precisas, capacitando al personal, monitoreando, poniendo los medios con debida diligencia, etc.? O, por el contrario, ¿puede ser penado aunque hubiera obrado bien? ¿Cuál es la responsabilidad de la persona jurídica? En un primer encuadre para responder a estas preguntas, partí de lo expresado por la Sala II de la Cámara Nacional de Apelaciones en lo Contencioso Administrativo Federal, que ha dicho que “la simple constatación de la comisión de infracciones genera la consiguiente responsabilidad y sanción del infractor, salvo que éste invoque y demuestre la existencia de una circunstancia exculpatoria válida”

Effects of seminal plasma and different cryoprotectants on rabbit sperm preservation at 16°c

The purpose of this research was to assess whether the presence of seminal plasma (SP) can improve sperm quality of rabbit spermatozoa stored at 16°C for 72 h and moreover evaluate the cryoprotectant effects of glycerol, N-N-Dimethylformamide (DMF), and N-methyl-2-pyrrolidone (NMP). Semen samples were pooled and divided in eight fractions. Four of them were diluted with INRA (extender A), INRA with 6% glycerol (extender B), INRA with 6% DMF (extender C), or INRA with 6% NMP (extender D), respectively. The other four fractions were centrifuged, and the supernatant was removed in order to eliminate SP. Each sample was then resuspended with extender A, B, C, or D, respectively. All samples were stored at 16°C and analysed at 4, 24, 48, and 72 h by ISAS®, vitality test, HOS test, and acrosome integrity test. After analyse of the results, SP samples showed a significantly higher percentage (P=0.020) in the HOS test (71.9 ± 1.6%) than non-SP samples (66.5 ± 1.6%). Non-SP samples had better results for kinematic parameters. Extenders A and C showed great results for the percentage of motile spermatozoa (63.1 ± 4.3% and 63.4 ± 3.7%, respectively), vitality (88.9 ± 2.6% and 87.7 ± 2.7%, respectively), and HOS test (68.9 ± 1.4% and 75.2 ± 1.4%, respectively). Extenders B and D showed worse data for sperm quality. These results suggest that SP has a protective effect on rabbit sperm membranes and maintains better sperm motility. The addition of glycerol and NMP to INRA does not improve rabbit sperm quality; nevertheless, the DMF cryoprotectant exerts a protective effect on the membrane of spermatozoa, improving seminal quality during rabbit sperm preservation at 16°C

Contribución al estudio de las esclerosis del páncreas en las gastroenteritis crónicas de los niños : Tesis presentada para optar al grado de doctor en medicina

A la cabeza de portada: Universidad Nacional de Buenos Aires. Facultad de Ciencias Médicas. - Incluye nómina de Catedráticos y Asignaturas. Tesis con dedicatoria