[National competitive bidding analysis of osteosynthesis materials in pediatric patients with femur fractures]. / Análisis de licitación pública nacional de materiales de osteosíntesis en los pacientes pediátricos con fracturas de fémur.

INTRODUCTION: health promotion policy requires the identification of barriers to the adoption of public policies. Paraguay's national healthcare system is inequitable, expensive, and inefficient. The Ministry of Public Health and Social Welfare (MSPyBS) is the entity responsible for covering the needs of a significant portion of the population. In January 2022, the MSPyBS financed the purchase of titanium elastic nails through a National Public Tender for Osteosynthesis Materials (LPN 02/22) to provide them for free in the pediatric service. Using research as a tool, we seek to analyze the impact of the implementation of LPN 02/22 at the Trauma Hospital, believing that this action would help streamline administrative and bureaucratic processes, making them more efficient with the assistance of the hospital's human resources. MATERIAL AND METHODS: a retrospective, analytical, and comparative study conducted at a high-complexity trauma center in Asunción, Paraguay. Patients aged 4 to 14 years with an indication for stabilization with elastic nails were included. Demographic data, the mechanism of injury, time elapsed from hospital arrival to surgical treatment, length of hospital stay, and the average hospital cost were analyzed based on the daily expense of pediatric patient hospitalization. RESULTS: 52 patients, divided into 25 cases in 2021 before implementation and 27 cases after implementation. The time elapsed from hospital arrival to definitive treatment was six days in the pre-implementation period, with an average stay from admission to discharge of 7.4 days. After implementation, the time from hospital arrival to definitive treatment was 4.3 days, and the average discharge time for the Post group was six days. The potential savings per patient amount to 332 dollars, offset by the institution's implant supply cost of 197 dollars, resulting in an approximate savings of 135 dollars per patient for the ministry. CONCLUSIONS: we view the implementation of free titanium elastic nails for pediatric femur fracture patients positively. We encourage the institution to continue with similar policies and strive to achieve even greater benefits for users.

INTRODUCCIÓN: la política de promoción de la salud requiere la identificación de los obstáculos para la adopción de políticas públicas. El sistema nacional de salud de Paraguay es inequitativo, caro e ineficiente. El Ministerio de Salud Pública y Bienestar Social (MSPyBS) es el ente que cubre las necesidades de gran parte de la población. El MSPyBS en Enero del 2022 financió, mediante la Licitación Pública Nacional de Materiales de Osteosíntesis (LPN 02/22), la compra de clavos elásticos de titanio para disponer de su uso gratuito en el Servicio de Pediatría; usando a la investigación como herramienta, buscamos analizar el impacto de la implementación de la LPN 02/22 en el Hospital de Trauma, creyendo que esta acción ayudaría a dinamizar los procesos administrativos y burocráticos, haciéndolos más eficientes con la ayuda de los recursos humanos del hospital. MATERIAL Y MÉTODOS: estudio retrospectivo, analítico y comparativo, realizado en un centro de trauma de alta complejidad de Asunción, Paraguay. Fueron incluidos los pacientes con edad comprendidas entre cuatro y 14 años, con indicación de estabilización con clavos elásticos. Se analizaron los datos demográficos, el mecanismo de trauma, el tiempo transcurrido desde la llegada al hospital hasta el tratamiento quirúrgico, así como el tiempo de estadía hospitalaria. Se evaluó el costo hospitalario promedio, basados en el gasto diario de la internación de un paciente pediátrico. RESULTADOS: cincuenta y dos pacientes, separados en 25 casos en el 2021 previo a la implementación y 27 casos posterior a la implementación. El tiempo transcurrido desde la llegada al hospital hasta el tratamiento definitivo fue de seis días para la etapa previa a la implementación; el promedio desde el ingreso hasta el alta fue de 7.4 días. Desde la implementación se tuvo un transcurso de 4.3 días desde la llegada al hospital hasta el tratamiento definitivo. El egreso del grupo Post tuvo un promedio de seis días. El ahorro probable en relación con cada paciente es de 332 dólares; a esto debemos contrarrestar el monto que paga la institución para la provisión del implante (197 dólares), por lo que el ahorro del ministerio sería de aproximadamente 135 dólares por cada paciente. CONCLUSIONES: vemos como positiva la implementación de la gratuidad de los clavos elásticos de titanio en los pacientes en edad pediátrica con fractura de fémur. Alentamos a la institución a seguir con políticas similares y tratar de lograr mayores beneficios para los usuarios.

Emerging challenges and opportunities in innovating food science technology and engineering education.

Progress in science, technology, innovation, and digital capabilities call for reassessing food science, technology, and engineering (FST&E) education and research programs. This survey targeted global professionals and students across food disciplines and nutrition. Its main objectives included assessing the status of FST&E higher education, identifying challenges and opportunities, and furnishing recommendations. Seven topics affecting the future of the FST&E curricula were evaluated by the panel as 'High' to 'Very high', namely: 'Critical thinking', followed by 'Problem-solving projects', 'Teamwork/collaboration', 'Innovation/Open innovation' and 'Multidisciplinary'. The importance of academic partnership/collaboration with the Food Industry and Nutrition Sciences was demonstrated. Significant positive roles of the food industry in collaboration and partnerships were found. Other essential food industry attributes were related to internships, education, strategy, and vision. Collaboration between FST&E and nutrition sciences indicated the high standing of this direction. The need to integrate or converge nutrition sciences and FST&E is emphasized, especially with the growing consumer awareness of health and wellness. The study provides insights into new education and learning opportunities and new topics for future curricula.

Accumulation and decomposition of cultural residues of Theobroma grandiflorum, Paullinia cupana, Bixa orellana and forest in the southern region of Amazonas.

The litter deposited on the soil surface at various stages of decomposition is important for primary productivity that impacts the microbial communities and soil carbon storage. The objective of this study was to evaluate the accumulation and decomposition of cultural residues of Theobroma grandiflorum (Willd. ex. Spreng) Schum, Paullinia cupana (Mart.) Ducke, Bixa orellana L., and forest in the Amazon region. The study was carried out in the São Francisco settlement, Canutama in the south of Amazonas, in a randomized block experimental design, and the treatments consisted of four areas with different crops: 1 - P. cupana; 2 - T. grandiflorum; 3 - B. orellana; 4 - Native woodland area (forest), in time subdivided plots: 7, 15, 30, 60, 90, 120, 150, 180, 210, 240, 270, 300, and 330 days after the distribution of the bags in the field, all with four repetitions. To evaluate the contribution and fractions of litter, conical collectors were used in each area, and collections were performed monthly in the period from March 2020 to February 2021. The estimate of the decomposition rate of the litter was done by quantifying the loss of mass, using litter bags, which allow for a direct analysis of the rate of decay over time. The forest and P. cupana environments presented the highest litter production, and greater deposition when compared to environments cultivated with T. grandiflorum and B. orellana. The forest and B. orellana areas showed the highest speed of decomposition, while the opposite situation occurred under T. grandiflorum and P. cupana cultivation.

The rs2165241 polymorphism of the Loxl1 gene in postmenopausal women with pelvic organ prolapse.

OBJECTIVE: This study aimed to verify the presence of polymorphism rs2165241 of the lysyl oxidase-like 1 (Loxl1) gene and its association with pelvic organ prolapse (POP) in Brazilian women and determine risk factors for POP development. METHODS: The study was previously approved by the local research and ethics board. Postmenopausal women were included and divided into POP (stages III and IV) and control (stages 0 and I) groups. Peripheral blood samples were collected, and the DNA sequence of interest was analyzed by real-time reverse-transcriptase polymerase chain reaction. We used logistic regression and considered a recessive model of inheritance for the analysis, with p < 0.05 for significance. RESULTS: A total of 836 women were assessed: 426 POP cases and 410 controls. The frequencies of CC, CT and TT genotypes were similar in both groups. Age (odds ratio [OR] = 1.1, 95% confidence interval [CI] = 1.07; 1.14), number of vaginal births (OR = 17.06, 95% CI = 5.94; 48.97), family history (OR = 2.87, 95% CI = 1.57; 5.22) and weight of largest newborn (OR = 1.001, 95% CI = 1.0003; 1.001) were independent risk factors for POP, while multiple cesarean sections (two or more) was protective (OR = 0.17, 95% CI = 0.07; 0.42). CONCLUSION: No association was detected between rs2165241 of the Loxl1 gene and POP.

Brain teratoma in a free-ranging mallard (Anas platyrhynchos) - case report / [Teratoma cerebral em um pato-real (Anas platyrhynchos) de vida livre - relato de caso]

Teratoma is a rare neoplasia with differentiation in two or three germ cell lines. Intracranial teratoma in birds has rarely been reported, especially affecting the brain. This is the first report of a brain teratoma in a mallard with neurological clinical signs. The neoplasm was characterized as a mature brain teratoma, extending from the cerebellum to the brainstem, and with one nodule in the cortex.(AU)

O teratoma é uma neoplasia rara, com diferenciação em duas ou três linhagens de células germinativas. Teratomas intracranianos, em aves, são raramente relatados, principalmente, com localização cerebral. Este é o primeiro relato de teratoma cerebral em um pato-real com sinais clínicos neurológicos. A neoplasia foi caracterizada como um teratoma cerebral maduro, estendendo do cerebelo ao tronco encefálico e com um nódulo no córtex.(AU)

Oral administration of Hsp65-producing Lactococcus lactis attenuates allergic asthma in a murine model.

AIMS: Allergic asthma is a chronic inflammatory lung disease characterized by a Th2-type immune response pattern. The development of nonspecific immunotherapy is one of the primary goals for the control of this disease. METHODS AND RESULTS: In this study, we evaluated the therapeutic effects of Lactococcus lactis-producing mycobacterial heat shock protein 65 (LLHsp65) in an ovalbumin (OVA)-induced allergic asthma model. OVA-challenged BALB/c mice were orally administrated with LLHsp65 for 10 consecutive days. The results demonstrate that LLhsp65 attenuates critical features of allergic inflammation, like airway hyperresponsiveness and mucus production. Likewise, the treatment decreases the pulmonary eosinophilia and the serum level of OVA-specific IgE. In addition to deviating immune responses towards Th1-cytokine profile, increase regulatory T cells, and cytokine levels, such as IL-6 and IL-10. CONCLUSIONS: Our results reveal that the mucosal immunotherapy of LLHsp65 significantly reduces the overall burden of airway allergic inflammation, suggesting a promising therapeutic strategy for allergic asthma treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: This research reveals new perspectives on nonspecific immunotherapy based on the delivery of recombinant proteins by lactic acid bacteria to treat of allergic disorders.

Efeito do hipotireoidismo materno na expressão espaço-temporal de mediadores imunológicos e população de células natural killers na decídua e na glândula metrial de ratas / Effect of maternal hypothyroidism on the spatiotemporal expression of immunological mediators and population of Natural Killers cells in decidua and metrial gland of rats

Estudou-se o efeito do hipotireoidismo materno na expressão espaço-temporal de mediadores imunológicos e na população de células natural killers (NK) na decídua e na glândula metrial de ratas durante a gestação. Avaliou-se a detecção imunoistoquímica de interferon γ (IFNγ), do fator inibidor de migração (MIF), da interleucina 15 (IL15), do óxido nítrico sintase induzível (iNOS), a marcação com lectina DBA para evidenciação das células NK uterinas DBA+ e a expressão gênica de Ifnγ e Nos2. O hipotireoidismo aumentou o iNOS aos sete dias, a IL15 e o MIF aos 10 e 12 dias, o IFNγ e o MIF aos 14 DG e a expressão dos transcritos gênicos para iNos aos 12 e 19 dias e para Ifnγ aos 14 DG. O hipotireoidismo reduziu a imunomarcação de MIF e lectina DBA aos sete dias, lectina DBA aos 10 e 14 DG, IFNγ aos 12 dias, e a expressão de Ifnγ aos 10 e 19 DG e de iNOS aos 12, 14 e 19 DG, bem como reduziu seus transcritos gênicos aos 10 e 14 DG. Conclui-se que o hipotireoidismo compromete o perfil imunológico na interface materno-fetal ao longo da gestação, particularmente por reduzir o fator anti-inflamatório iNOS e a população de células uNK DBA+.(AU)

The goal of this study was to evaluate the effect of maternal hypothyroidism on the spatiotemporal expression of immunological mediators and population of Natural Killers cells in decidua and metrial gland of rats. Interferon gamma (IFNγ), migration inhibitory factor (MIF), interleukin 15 (IL15), inducible nitric oxide synthase (iNOS), and DBA-Lectin labeling for evidence of uNK DBA+ cells in decidua and genetic expression of Ifnγ and iNos by real-time RT-PCR were evaluated. Hypothyroidism increased protein expression of iNOS at 7 days, IL15 and MIF at 10 and 12 days, IFNγ and MIF at 14 DG in the decidua and/or metrial gland and the gene transcripts for iNOS at 12 and 19 days and for Inf at 14 DG. In addition, hypothyroidism reduced the protein expression of MIF and DBA-Lectin at 7 days, DBA-Lectin at 10 and 14 DG, IFNγ at 12 days, and the gene transcript to Ifnγ at 10 and 19 DGs. Hypothyroidism also reduced the protein expression of iNOS at 12, 14 and 19 DG and reduced its gene transcripts at 10 and 14 DGs. It is concluded that hypothyroidism compromises the immunology profile at the maternal-fetal interface throughout pregnancy, particularly by reducing the anti-inflammatory factor iNOS and population of uNK DBA+ cells.(AU)

Norovirus recovery from floors and air after various decontamination protocols.

BACKGROUND: The dispersal of airborne norovirus (NoV) particles from the floor after contamination with faeces or vomit is a challenge for infection control, as this pathogen is infectious at low doses. Therefore, it is imperative to establish a safe protocol for floor decontamination. AIM: To assess the presence of residual NoV-GII particles on floors and airborne particles following various floor decontamination procedures. METHODS: Two types of floor (vinyl and granite) were contaminated intentionally with 10% human faeces, positive for NoV-GII. Two decontamination protocols were implemented: cleaning followed by disinfection using 1% sodium hypochlorite, and cleaning followed by disinfection using a manual ultraviolet C (UV-C) light device. Swab samples were taken from the floors, and air samples were obtained using an air sampler. The TaqMan method for real-time reverse transcription-quantitative polymerase chain reaction was employed for analysis. FINDINGS: The disinfection protocol using 1% sodium hypochlorite after cleaning proved to be more effective than cleaning followed by UV-C light exposure (P<0.001). Viral particles were detected in 27 of 36 air samples after cleaning, with no significant difference between the two floor types. On average, 617 genome copies/sample were identified in air samples after cleaning, but the number decreased gradually after disinfection. CONCLUSION: NoV-GII can be aerosolized during floor cleaning, and its particles may be inhaled and then swallowed or can settle on surfaces. Therefore, residual viral particles on floors must be fully eliminated. Cleaning followed by 10 min of 1% sodium hypochlorite disinfection proved to be the superior decontamination protocol.

CCAAT/enhancer-binding protein alpha (CEBPA) gene haploinsufficiency does not alter hematopoiesis or induce leukemia in Lck-CALM/AF10 transgenic mice.

Although rare, CALM/AF10 is a chromosomal rearrangement found in immature T-cell acute lymphoblastic leukemia (T-ALL), acute myeloid leukemia, and mixed phenotype acute leukemia of T/myeloid lineages with poor prognosis. Moreover, this translocation is detected in 50% of T-ALL patients with gamma/delta T cell receptor rearrangement, frequently associated with low expression of transcription factor CCAAT/enhancer-binding protein alpha (CEBPA). However, the relevance of CEBPA low expression for CALM/AF10 leukemogenesis has not yet been evaluated. We generated double mutant mice, which express the Lck-CALM/AF10 fusion gene and are haploinsufficient for the Cebpa gene. To characterize the hematopoiesis, we quantified hematopoietic stem cells, myeloid progenitor cells, megakaryocyte-erythrocyte progenitor cells, common myeloid progenitor cells, and granulocyte-macrophage progenitor cells. No significant difference was detected in any of the progenitor subsets. Finally, we tested if Cebpa haploinsufficiency would lead to the expansion of Mac-1+/B220+/c-Kit+ cells proposed as the CALM/AF10 leukemic progenitor. Less than 1% of bone marrow cells expressed Mac-1, B220, and c-Kit with no significant difference between groups. Our results showed that the reduction of Cebpa gene expression in Lck-CALM/AF10 mice did not affect their hematopoiesis or induce leukemia. Our data corroborated previous studies suggesting that the CALM/AF10 leukemia-initiating cells are early progenitors with lymphoid/myeloid differentiating potential.

CCAAT/enhancer-binding protein alpha (CEBPA) gene haploinsufficiency does not alter hematopoiesis or induce leukemia in Lck-CALM/AF10 transgenic mice

Although rare, CALM/AF10 is a chromosomal rearrangement found in immature T-cell acute lymphoblastic leukemia (T-ALL), acute myeloid leukemia, and mixed phenotype acute leukemia of T/myeloid lineages with poor prognosis. Moreover, this translocation is detected in 50% of T-ALL patients with gamma/delta T cell receptor rearrangement, frequently associated with low expression of transcription factor CCAAT/enhancer-binding protein alpha (CEBPA). However, the relevance of CEBPA low expression for CALM/AF10 leukemogenesis has not yet been evaluated. We generated double mutant mice, which express the Lck-CALM/AF10 fusion gene and are haploinsufficient for the Cebpa gene. To characterize the hematopoiesis, we quantified hematopoietic stem cells, myeloid progenitor cells, megakaryocyte-erythrocyte progenitor cells, common myeloid progenitor cells, and granulocyte-macrophage progenitor cells. No significant difference was detected in any of the progenitor subsets. Finally, we tested if Cebpa haploinsufficiency would lead to the expansion of Mac-1+/B220+/c-Kit+ cells proposed as the CALM/AF10 leukemic progenitor. Less than 1% of bone marrow cells expressed Mac-1, B220, and c-Kit with no significant difference between groups. Our results showed that the reduction of Cebpa gene expression in Lck-CALM/AF10 mice did not affect their hematopoiesis or induce leukemia. Our data corroborated previous studies suggesting that the CALM/AF10 leukemia-initiating cells are early progenitors with lymphoid/myeloid differentiating potential.

Thermodynamics of the adsorption of monoclonal antibodies in phenylboronate chromatography: Affinity versus multimodal interactions.

The aim of this work was to investigate the complex phenomena underlying the adsorption of an anti-human IL-8 (anti-IL8) monoclonal antibody (mAb) to m-aminophenylboronate (m-APBA) by Flow Microcalorimetry (FMC) and to understand the role of non-specific interactions in the adsorption process. FMC was exploited as a dynamic on-line method to measure instantaneous heat energy transfers in order to understand the surface phenomena underlying mAb's adsorption towards the synthetic ligand m-APBA under different pH (7.5, 8.5, 9.0, 9.5 and 10.0) and salt concentrations (0 and 150 mM NaCl). Results showed that the adsorption of anti-IL8 mAb to m-APBA is enthalpically driven (ΔHads<0), as expected for the predominant reversible esterification reaction between boronates and cis-diols-containing molecules. For all the pH conditions studied, thermograms presented a first exothermic peak, characteristic of the reversible esterification reaction between mAb (pI≥9.3) and m-APBA (pKa = 8.8), except at pH 9.0 in the presence of 150 mM NaCl, for which the thermogram presented a first endothermic peak. The heat of adsorption (ΔHads) obtained at conditions where cis-diol interactions were predominant was approximately -243 ± 38 kJ/mol against -82 ± 14 kJ/mol (p-value < 0.05) obtained at pH 9.0 with 150 mM NaCl. The observed shift in the thermogram profile at pH 9.0, 150 mM NaCl, and the consequent decrease of 60-70% in ΔHads were indicative of the promotion of electrostatic interactions between the protein and the ligand. Overall, and whereas the binding of the PBA ligand to mAb molecules has been described for decades as being affinity-based, our study demonstrates the multimodal behaviour of this interaction and contributes towards the understanding of the adsorption thermodynamics.

Value of the oxygen pulse curve for the diagnosis of coronary artery disease.

This study investigated the value of oxygen (O(2)) pulse curves obtained during cardiopulmonary exercise testing (CPET) for the diagnosis of coronary artery disease (CAD). Forty patients with known coronary anatomy (35.0 % normal, 27.0 % single-vessel and 38.0 % multivessel CAD) underwent CPET with radiotracer injection at peak exercise, followed by myocardial scintigraphy. O(2) pulse curves were classified as: A-normal, B-probably normal (normal slope with low peak value); C-probably abnormal (flat, with low peak value); or D-definitely abnormal (descending slope). Sensitivity, specificity, positive and negative predictive values of the O(2) pulse curve pattern (A or B vs. C or D) for the diagnosis of CAD were, respectively, 38.5 %, 81.3 %, 76.9 %, and 44.8 %. The concordance rate between the abnormal O(2) pulse curve pattern and ischemia in myocardial scintigraphy was 38.1 %. Age and the extent of scintigraphic perfusion defect, but not the abnormal O(2) pulse curve patterns (B or C or both combined) were independently associated with CAD. In conclusion, the O(2) pulse curve pattern has low diagnostic performance for the diagnosis of obstructive CAD, and the abnormal curve pattern was not associated with myocardial ischemia defined by scintigraphy.

Resistance in 27 Rice Cultivars to Sugarcane Borer (Lepidoptera: Crambidae).

Rice (Oryza sativa L.) is considered one of the most important crops in the world, and the sugarcane borer, Diatraea saccharalis Fabricius (Lepidoptera: Crambidae), is among the key pests damaging the crop in the Americas. The objective of this work was to identify rice genotypes as a source of resistance to D. saccharalis. Rice plants were infested in the greenhouse and subsequently evaluated for damage, larval weight and survival, and stem size. The cultivars 'Bonança', 'Caripuna', 'IR 42', 'Canela de Ferro', 'SWA Norte', 'BR IRGA 409', 'Pepita', 'Serra Dourada', 'Araguaia', 'Xingú', 'Tangará', and 'Soberana' showed antibiosis antixenosis, or both to D. saccharalis. These cultivars may be used as donor sources in the breeding program and used directly by Brazilian farmers as a component of rice-integrated pest management.

Osteogenic differentiation of adipose tissue-derived mesenchymal stem cells cultured with different concentrations of prolactin / Diferenciação osteogênica de células tronco mesenquimais do tecido adiposo cultivadas com diferentes concentrações de prolactina

The objective was to evaluate the in vitro effect of prolactin in osteogenic potential of adipose tissue-derived mesenchymal stem cells (ADSCs) in female rats. ADSCs were cultured in osteogenic medium with and without the addition of prolactin and distributed into three groups: 1) ADSCs (control), 2) ADSCs with addition of 100ng/mL of prolactin and 3) ADSCs with addition of 300ng/mL of prolactin. At 21 days of differentiation, the tests of MTT conversion into formazan crystals, percentage of mineralized nodules and cells per field and quantification of genic transcript for alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I by real-time RT-PCR were made. The addition of prolactin reduced the conversion of MTT in group 3 and increased the percentage of cells per field in the groups 2 and 3, however without significantly increasing the percentage of mineralized nodules and the expression of alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I. In conclusion, the addition of prolactin in concentrations of 100ng/mL and 300ng/mL does not change the osteogenic differentiation to the ADSCs of female rats despite increase in the cellularity of the culture.(AU)

O objetivo do presente trabalho foi avaliar o efeito in vitro da prolactina sobre o potencial osteogênico de células-tronco mesenquimais do tecido adiposo (CTM-TA) em ratas. CTM-TA foram cultivadas em meio osteogênico com e sem adição de prolactina e distribuídas em três grupos: 1) CTM-TA (controle), 2) CM-TA com adição de 100ng/mL de prolactina e 3) CTM-TA com adição de 300ng/mL de prolactina. Aos 21 dias de diferenciação, foram realizados os testes de conversão do MTT em cristais de formazan, porcentagem de nódulos mineralizados e células por campo e quantificação dos transcritos gênicos para fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. A adição de prolactina reduziu a conversão do MTT no grupo 3 e aumentou a porcentagem de células por campo nos grupos 2 e 3, sem alterar significativamente a porcentagem de nódulos mineralizados e a expressão de fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. Conclui-se que a adição de prolactina nas concentrações de 100ng/mL e 300ng/mL não altera a diferenciação osteogênica das CTM-TA de ratas, apesar do aumento de celularidade da cultura.(AU)

Relationship between ovulation rate and embryonic characteristics in gilts at 35 d of pregnancy.

This study investigated the relationship between ovulation rate (OR) and embryonic characteristics in gilts. Landrace ( = 86) and Yorkshire x Landrace ( = 212) gilts were inseminated with semen stored for 3 to 5 d (SS1, = 59), 6 to7 d (SS2, = 133), or 8 to 10 d (SS3, = 106), and slaughtered at 35 d of pregnancy. Ovulation rate was assessed by dissection of the corpora lutea on both ovaries. Embryos were classified as vital (VE) by visual appearance and individually weighed (VEg) and the SD of the weight calculated (SDVEg). Early embryonic mortality (EM) was estimated as the difference between OR and the number of vital plus nonvital embryos. Embryonic characteristics were analyzed with a model that included linear and quadratic terms of OR and fixed class effects of semen storage duration (SS) and genetic line (GL). Landrace gilts had a higher OR than Yorkshire x Landrace gilts (22.1 ± 0.4 vs. 20.3 ± 0.2, ≤ 0.05) and also a higher EM (6.1 ± 0.4 vs. 3.5 ± 0.3, ≤ 0.05). EM was also higher in gilts inseminated with semen stored for more than 8 d. Also, Yorkshire x Landrace gilts had a higher number of VE (16.9 ± 0.7) than the Landrace gilts (13.3 ± 0.8) when inseminations were done with semen stored for up to 5 d. Yorkshire x Landrace gilts had the highest VEg when inseminated with semen stored for 3 to 5 d (SS1: 4.9 ± 0.2 g, SS2: 4.1 ± 0.1 g, and SS3: 4.0 ± 0.2 g; ≤ 0.05). VE and VEg did not differ within Landrace gilts between different SS classes. A quadratic relationship of OR ( ≤ 0.05) was found with VE: a maximum of 16.8 VE was observed at 26 ovulations [(2.5 (± 0.6)*OR- 0.05 (± 0.01)*OR]. A quadratic relationship of OR ( ≤ 0.05) was also found for EM: a minimum of 3.33 EM was observed at 15 ovulations [(-1.1 (± 0.6)*OR -0.03 (± 0.01)*OR]. VEg was not related with OR, but SDVEg had a positive linear relationship with OR [0.01 (± 0.003)*OR, ≤ 0.05]. Results show that Yorkshire x Landrace gilts perform better than Landrace when inseminated with fresh semen, but not with semen stored for longer time. Also, the VE increases with an increase in OR up to 26, but at a lower level at higher OR, which is likely related with the increase in EM. The higher EM at higher OR might arise from a higher variation in follicular/oocyte quality leading to a higher variation in embryonic quality and development, increasing mortality before uterine implantation and the variation in embryonic weight already at 35 d of pregnancy.

Validation of transrectal ultrasonography for assessment of corpora lutea characteristics in pregnant sows and its relationship with litter characteristics at birth.

In experiment 1 we investigated the accuracy of transrectal ultrasonography (TUS) to assess the number (OR) and diameter of corpora lutea (CL) in 45 and 25 sows, respectively, at 23.4 ± 2.9 d of pregnancy. The diameter was calculated as the average diameter of 10 biggest CL. Sows were subsequently slaughtered and OR was assessed by dissection of CL from both ovaries (n = 45) and average diameter of the 10 biggest CL was also calculated after measurement of CL with the caliper rule (n = 25). There was a weak relationship between OR counted after dissection of the ovaries and OR counted with TUS (ß = 0.28 ± 0.01 CL/CL, P = 0.01), but there was a strong relationship between the average CL diameter measured with the caliper rule after dissection and the average CL diameter based on TUS (ß = 1.0 ± 0.1 mm/mm, P < 0.0001). This shows that TUS is not a valid method to assess OR in pregnant sows but it is a valid method to assess average CL diameter. In experiment 2, we investigated the relationship between the average CL diameter assessed by TUS (n = 100) at 23.8 ± 2.4 d of pregnancy and average piglet birth weight (BW) and observed an increase of 37.6 ± 17.8 g in piglet BW per mm increase in average CL diameter measured by TUS (P = 0.04). This relationship is probably because larger CL develop from bigger follicles at ovulation, which might have ovulated oocytes of higher quality that developed into embryos with higher growth potential and thus higher birth weight.

ADME studies and preliminary safety pharmacology of LDT5, a lead compound for the treatment of benign prostatic hyperplasia.

This study aimed to estimate the absorption, distribution, metabolism and excretion (ADME) properties and safety of LDT5, a lead compound for oral treatment of benign prostatic hyperplasia that has previously been characterized as a multi-target antagonist of α1A-, α1D-adrenoceptors and 5-HT1A receptors. The preclinical characterization of this compound comprised the evaluation of its in vitro properties, including plasma, microsomal and hepatocytes stability, cytochrome P450 metabolism and inhibition, plasma protein binding, and permeability using MDCK-MDR1 cells. De-risking and preliminary safety pharmacology assays were performed through screening of 44 off-target receptors and in vivo tests in mice (rota-rod and single dose toxicity). LDT5 is stable in rat and human plasma, human liver microsomes and hepatocytes, but unstable in rat liver microsomes and hepatocytes (half-life of 11 min). LDT5 is highly permeable across the MDCK-MDR1 monolayer (Papp ∼32×10-6 cm/s), indicating good intestinal absorption and putative brain penetration. LDT5 is not extensively protein-bound and is a substrate of human CYP2D6 and CYP2C19 but not of CYP3A4 (half-life >60 min), and did not significantly influence the activities of any of the human cytochrome P450 isoforms screened. LDT5 was considered safe albeit new studies are necessary to rule out putative central adverse effects through D2, 5-HT1A and 5-HT2B receptors, after chronic use. This work highlights the drug-likeness properties of LDT5 and supports its further preclinical development.

Optimizing the transport of porcine semen: a proposal for Brazil / Otimizar o transporte do sêmen suíno: uma proposta para o Brasil

Semen from the first 15mL of the ejaculate (P1) obtained from two boars (30mL) was diluted in glycine-egg yolk extender, cooled at 5°C in a special container and rediluted in standard doses of 3x109 mobile spermatozoa after 12h of storage. Semen was also stored up to 24h after redilution. The physical characteristics of the semen were evaluated at different storage periods (fresh, 0h, 12h, rediluted, 24h, and 36h). The reproductive performance of the boars and their fertility regarding the insemination of primiparous sows were also determined. Two treatments were used: T1-15B sows inseminated with semen originated from hyperconcentrated heterospermic doses (15x109 mobile spermatozoa per dose), rediluted after 12h of storage at 5°C for standard doses of 3x109 mobile spermatozoa per dose and stored at 5°C up to 24h after redilution (n=10); T2-3B sows inseminated with standard heterospermic doses (3x109 mobile spermatozoa per dose), stored at 5°C up to 36h after semen collection (n=10). There was no effect (P>0.05) of treatments on the spermatic motility, even though a pronounced decrease (P>0.05) of their values at 12h of storage was recorded. However, they remained higher than 70% until 36h. There was effect of treatments on spermatic vigour at 0h (P<0.05), when T1-15B vigour was higher. There was also effect of the storage period for both treatments with a progressive decrease throughout 36h of storage, although the differences were not always significant. Pregnancy rates (90%) and the number of total farrowed piglets (15, 11-T1-15B; 13, 44- T2-3B) did not differ (P>0.05) between the treatments. It was concluded that the semen hyperconcentration of 15 billion of mobile spermatozoa per dose, stored at 5°C for 12h, did not result in drawbacks considering the physical characteristics of the semen, maintaining the pregnancy rates and prolificacy of the inseminated sows.

Os primeiros 15mL do ejaculado (P1) de dois varrões foram coletados (30mL) e diluídos em diluidor glicina-gema de ovo, resfriados a 5°C em contêiner especial e rediluídos para doses padrão de 3x109 espermatozoides (sptz) móveis, após 12 horas de armazenamento. Além disso, foram armazenados por até 24 horas após a rediluição, sendo as características físicas avaliadas em diferentes períodos de estocagem (fresco, zero hora, 12h, Red12h, 24h e 36h) e a fertilidade avaliada por meio de fêmeas primíparas inseminadas. Foram realizados dois tratamentos: T1-15B: porcas inseminadas com sêmen de doses heterospérmicas hiperconcentradas (15x109 sptz móveis/dose), rediluídas após 12 horas de armazenamento a 5°C para doses padrão de 3x109 sptz móveis/dose, e armazenadas a 5°C por até 24 horas após a rediluição (n=10); T2-3B: porcas inseminadas com doses heterospérmicas padrão (3x109 sptz móveis/dose), armazenadas a 5°C por até 36 horas após coleta. Não houve efeito (P>0.05) dos tratamentos sobre a motilidade espermática e, embora tenha ocorrido queda (P<0.05) às 12 horas, a motilidade foi superior a 70% durante as 36 horas de armazenamento. Houve efeito (P<0.05) dos tratamentos no tempo zero hora quanto ao vigor espermático, sendo E1T1-15B superior. Além disso, houve efeito do período de estocagem para os dois tratamentos, com queda progressiva do vigor ao longo das 36 horas, embora nem sempre as diferenças tenham sido significativas. As taxas de gestação (90%) e o número total de leitões nascidos (15, 11 - T1-15B; 13, 44 - T2-3B) não diferiram (P>0.05) entre os tratamentos. Concluiu-se que a hiperconcentração do sêmen para 15x109 sptz móveis/dose, armazenado a 5°C por 12 horas não resultou em prejuízos quanto à manutenção das características físicas do sêmen e ao desempenho reprodutivo dos varrões, sendo capaz de manter a taxa de gestação e a prolificidade das fêmeas inseminadas.

Extracellular matrix microarrays to study inductive signaling for endoderm specification.

During tissue development, stem and progenitor cells are faced with fate decisions coordinated by microenvironmental cues. Although insights have been gained from in vitro and in vivo studies, the role of the microenvironment remains poorly understood due to the inability to systematically explore combinations of stimuli at a large scale. To overcome such restrictions, we implemented an extracellular matrix (ECM) array platform that facilitates the study of 741 distinct combinations of 38 different ECM components in a systematic, unbiased and high-throughput manner. Using embryonic stem cells as a model system, we derived definitive endoderm progenitors and applied them to the array platform to study the influence of ECM, including the interactions of ECM with growth factor signaling, on the specification of definitive endoderm cells towards the liver and pancreas fates. We identified ECM combinations that influence endoderm fate decisions towards these lineages, and demonstrated the utility of this platform for studying ECM-mediated modifications to signal activation during liver specification. In particular, defined combinations of fibronectin and laminin isoforms, as well as combinations of distinct collagen subtypes, were shown to influence SMAD pathway activation and the degree of hepatic differentiation. Overall, our systematic high-throughput approach suggests that ECM components of the microenvironment have modulatory effects on endoderm differentiation, including effects on lineage fate choice and cell adhesion and survival during the differentiation process. This platform represents a robust tool for analyzing effects of ECM composition towards the continued improvement of stem cell differentiation protocols and further elucidation of tissue development processes. STATEMENT OF SIGNIFICANCE: Cellular microarrays can provide the capability to perform high-throughput investigations into the role of microenvironmental signals in a variety of cell functions. This study demonstrates the utility of a high-throughput cellular microarray approach for analyzing the effects of extracellular matrix (ECM) in liver and pancreas differentiation of endoderm progenitor cells. Despite an appreciation that ECM is likely involved in these processes, the influence of ECM, particularly combinations of matrix proteins, had not been systematically explored. In addition to the identification of relevant ECM compositions, this study illustrates the capability of the cellular microarray platform to be integrated with a diverse range of cell fate measurements, which could be broadly applied towards the investigation of cell fate regulation in other tissue development and disease contexts.