Dynamics of CD44+ bovine nucleus pulposus cells with inflammation.

Intervertebral Disc (IVD) degeneration has been associated with a chronic inflammatory response, but knowledge on the contribution of distinct IVD cells, namely CD44, to the progression of IVD degeneration remains elusive. Here, bovine nucleus pulposus (NP) CD44 cells were sorted and compared by gene expression and proteomics with the negative counterpart. NP cells were then stimulated with IL-1b (10 ng/ml) and dynamics of CD44 gene and protein expression was analyzed upon pro-inflammatory treatment. The results emphasize that CD44 has a multidimensional functional role in IVD metabolism, ECM synthesis and production of neuropermissive factors. CD44 widespread expression in NP was partially associated with CD14 and CD45, resulting in the identification of distinct cell subsets. In conclusion, this study points out CD44 and CD44-based cell subsets as relevant targets in the modulation of the IVD pro-inflammatory/degenerative cascade.

Magnesium incorporation in fibrinogen scaffolds promotes macrophage polarization towards M2 phenotype.

The host inflammatory response to biomaterials conditions their capacity to promote tissue repair, and macrophage polarization shift from M1 to M2 is determinant in this process. Previous work showed that extracts of a combination between fibrinogen and metallic magnesium materials acted synergistically to reduce macrophage inflammatory phenotype. The hypothesis underlying the current work was that the ability of magnesium-modified fibrinogen scaffolds to modulate macrophage phenotype depends on the concentration of magnesium. Thus, Fibrinogen (Fg) scaffolds incorporating precise concentrations of magnesium sulfate (Mg: 0, 10, 25, 50 mM) were developed and characterized. Mg incorporation in Fg scaffolds increased surface charge, while porosity decreased with increasing Mg concentrations, but only Fg scaffolds with 10 mM of Mg (FgMg10) had significantly improved mechanical properties. Human macrophages cultured on FgMg10 scaffolds, showed increased M2 and decreased M1 polarization, when compared to those cultured on scaffolds with 0, 25 and 50 mM of Mg. Macrophage polarization results were independent of the anion used (chloride or sulfate). Macrophage modulation by FgMg10 scaffolds involved reduced NF-κB p65 nuclear translocation, and impacted production of pro-inflammatory mediators (e.g. IFNγ, IL-12, TNF-⍺, IP-10). Importantly, FgMg10 scaffolds implanted in vivo increased the expression of M2 marker CD163, in macrophages from inflammatory exudates, compared to Sham and Fg-implanted animals, increasing the M2:M1 ratio. A cytokine/chemokine array showed that, while both Fg and FgMg10 scaffolds decreased inflammatory mediators, only FgMg10 decreased IL-1ß, IP-10, MIP-2, MDC and MIP-3⍺, compared to Sham-operated animals. This study demonstrated that incorporation of 10mM of Mg modulated inflammation, promoting M2 macrophage polarization in vitro and in vivo. STATEMENT OF SIGNIFICANCE: Developing biomaterials that can modulate inflammation and promote macrophage phenotype switch from M1 to M2 is crucial to promote a regenerative microenvironment. Our previous work showed that extracts of a combination between fibrinogen (Fg) and metallic magnesium (Mg) materials synergistically reduced macrophage pro-inflammatory phenotype. Herein, we tested the hypothesis that macrophage modulation was dependent on Mg concentration. A new family of Fg porous scaffolds incorporating different amounts of Mg (0, 10, 25 and 50 mM) was produced and characterized. We observed that only the combination of Fg scaffolds with 10 mM of Mg (FgMg10) significantly changed the scaffolds mechanical properties and directed macrophages towards a M2 phenotype, reducing the production of inflammatory mediators, both in vitro and in vivo.

Fibrotic alterations in human annulus fibrosus correlate with progression of intervertebral disc herniation.

BACKGROUND: Intervertebral disc (IVD) herniation is characterized by annulus fibrosus failure (AF) in containing the nucleus pulposus (NP). IVD herniation involves cellular and extracellular matrix (ECM) alterations that have been associated with tissue fibrosis, although still poorly investigated. METHODS: Here, fibrotic alterations in human AF were evaluated, by characterizing the herniated ECM. Human AF samples (herniated lumbar IVD (n = 39, age 24-83) and scoliosis controls (n = 6, age 15-21)) were processed for transmission electron microscopy and histological/immunohistochemical analysis of fibrotic markers. Correlations between the fibrotic markers in AF ECM and the degree of NP containment (protused, contained and uncontained) and patients' age were conducted. RESULTS: Our results demonstrate that with herniation progression, i.e. loss of NP containment, human AF presents less stained area of sulphated glycosaminoglycans and collagen I, being collagen I fibres thinner and disorganized. On the other hand, fibronectin stained area and percentage of α-smooth muscle actin+ cells increase in human AF, while matrix metalloproteinase-12 (MMP12) production and percentage of macrophages (CD68+ cells) remain constant. These structural and biochemical fibrotic alterations observed in human AF with herniation progression occur independently of the age. CONCLUSIONS: The characterization of human AF here conducted evidence the presence of fibrosis in degenerated IVD, while highlighting the importance of considering the herniation progression stage, despite the patients' age, for a better understanding of the mechanisms behind AF failure and IVD herniation.

IL-1ß-pre-conditioned mesenchymal stem/stromal cells' secretome modulates the inflammatory response and aggrecan deposition in intervertebral disc.

Mesenchymal stem/stromal cells (MSCs) have been increasingly used in clinical trials for low-back pain (LBP) and intervertebral disc (IVD) degeneration with promising results. Their action mechanisms are not fully understood, but they reduce IVD pro-inflammatory markers in a pro-inflammatory/degenerative IVD microenvironment. In this study the therapeutic potential of the MSC secretome, as an alternative cell-free approach for treating degenerated IVDs, was examined. Human bone marrow-derived MSC secretome (MSCsec) was collected after 48 h of preconditioning in IL-1ß (10 ng/mL) and low oxygen (6 % O2), mimicking the degenerative IVD. IL-1ß-pre-conditioning of MSCs increased secretion of pro-inflammatory markers hIL-6, hIL-8, hMCP-1, etc. The therapeutic effect of MSCsec was tested in a pro-inflammatory/degenerative IVD ex vivo model. MSCsec down-regulated IVD gene expression of pro-inflammatory cytokines (bIL-6, bIL-8) and matrix degrading enzyme bMMP1, while bMMP3 and bTIMP2 were up-regulated, at 48 h. After 14 d, MSCsec-treated IVDs revealed increased aggrecan deposition, although no differences in other ECM components were observed. Protein analysis of the MSCsec-treated IVD supernatant revealed a significant increase of CXCL1, MCP-1, MIP-3α, IL-6, IL-8 and GRO α/ß/γ (related to TNF, NOD-like receptor and neutrophil chemotaxis signalling), and a decrease of IFN-γ, IL-10, IL-4, IL-5 and TNF-α (associated with T-cell receptor signalling). MSCsec-treated IVD supernatants did not promote angiogenesis and neurogenesis in vitro. Overall, MSCsec can be a safe therapeutic approach, presenting a strong immunomodulatory role in degenerated IVD while potentiating aggrecan deposition, which can open new perspectives on the use of MSCsec as a cell-based/ cell-free therapeutic approach to LBP.

B-mode and Doppler ultrasonography of abdominal structures of the peccary (Tayassu tajacu, Linnaeus 1758) / [Ultrassonografias Doppler e modo-B das estruturas abdominais de catetos (Tayassu tajacu Linnaeus, 1758)]

This study used B-mode and Doppler ultrasonography to characterize the abdominal structures of healthy peccaries raised in captivity. Fifteen peccaries were used for this study. The urinary vesicle appeared as an ovoid structure, located in the abdominal and pelvic transition, with a hyperechogenic, thin, smooth, and regular wall. The kidneys presented retroperitoneal topography and had similar sizes. The kidney/aorta ratio had an average value of 10.53±15cm (right) and 10.23±0.12 (left). The right adrenal gland had a length of 1.93±0.34cm and diameter of 0.56±0.16cm. The left adrenal gland had a length of 1.85±0.42cm and diameter of 0.52±0.11cm. The spleen had a diameter of 1.13±0.18cm. The hepatic vein demonstrated polyphasic flow in pulsed Doppler, with two retrograde peaks and an anterograde peak with a flow velocity of 25.7±0.83cm/s. The abdominal aorta had a diameter of 0.58±0.05cm and a flow velocity of 115.17±5.32cm/s. The morphological and hemodynamic study of the abdominal structures of the peccary, observed through B-mode and Doppler ultrasonography, aided in identifying the size, shape, position, echogenicity, and echotexture of the abdominal organs and in making inferences about the normal parameters for these structures in this species.(AU)

Este estudo teve como objetivo utilizar as ultrassonografias de modo-B e Doppler para caracterizar as estruturas abdominais de um cateto sadio criado em cativeiro. Quinze catetos foram utilizados para este estudo. A vesícula urinária apareceu como uma estrutura ovoide, localizada na transição entre as partes abdominal e pélvica, com uma parede hiperecogênica, fina, lisa e regular. Os rins apresentaram topografia retroperitoneal e tamanhos semelhantes. A relação rim/aorta teve um valor médio de 10,53 ± 15cm (direita) e 10,23 ± 0,12cm (esquerda). A glândula adrenal direita tinha um comprimento de 1,93 ± 0,34cm e um diâmetro de 0,56 ± 0,16cm. A glândula suprarrenal esquerda tinha um comprimento de 1,85 ± 0,42cm e um diâmetro de 0,52 ± 0,11cm. O baço tinha um diâmetro de 1,13 ± 0,18cm. A veia hepática demonstrou fluxo polifásico no Doppler pulsátil, com dois picos retrógrados e um pico anterógrado com velocidade de fluxo de 25,7±0,83cm/s. A aorta abdominal tinha um diâmetro de 0,58 ± 0,05cm e uma velocidade de fluxo de 115,17±5,32cm/s. Os estudos morfológico e hemodinâmico das estruturas abdominais do queixada, observadas por meio das ultrassonografias modo-B e Doppler, auxiliaram na identificação do tamanho, da forma, da posição, da ecogenicidade e da ecotextura dos órgãos abdominais e na realização de inferências sobre os parâmetros de normalidade para as estruturas nas espécies.(AU)

Maternal separation induces changes in TREK-1 and 5HT1A expression in brain areas involved in the stress response in a sex-dependent way.

Depression is a prevalent disease in modern society, and has been linked to stressful events at early ages. Women are more susceptible to depression, and the neural basis for this are still under investigation. Serotonin is known to be involved in depression, and a decrease in 5HT1A expression is observed on temporal and cortical areas in both men and women with depression. As knockout animals for TREK-1 are resilient to depression, this channel has emerged as a new potential pharmacological target for depression treatment. In this study, maternal separation (MS) was used to emulate early-life stress, and evaluate behaviour, as well as TREK-1 and 5HT1A expression in the brain using immunohistochemistry. In juvenile females, 5HT1A reduction coupled to increased TREK-1 in the dentate gyrus (DG) was associated with behavioural despair, as well as increased TREK-1 expression in basolateral amygdala (BLA) and prelimbic cortex (PL). In juvenile males, MS induced an increase in 5HT1A in the BLA, and in TREK-1 in the PL, while no behavioural despair was observed. Anhedonia and anxiety-like behaviour were not induced by MS. We conclude stress-induced increase in TREK-1 in PL and GD is associated to depression, while 5HT1A changes coupled to TREK-1 changes may be necessary to induce depression, with females being more vulnerable to MS effects than males. Thus, TREK-1 and 5HT1A may be potential pharmacological targets for antidepressants development.

Role of Recombinant Parvalbumin Gad c 1 in the Diagnosis and Prognosis of Fish Allergy.

BACKGROUND AND OBJECTIVES: The prevalence of fish allergy has increased in recent years. The parvalbumin Gad c 1 is a major cod allergen that is used as a follow-up marker in patients with fish allergy. Objectives: To determine the clinical and laboratory characteristics of a population of patients with fish allergy. To analyze the role of the specific IgE (sIgE) of recombinant Gad c 1 (rGad c 1) and skin prick tests (SPTs) in confirming the acquisition of tolerance to fish. METHODS: We performed a retrospective study of patients with fish allergy from July 1, 2005 to December 31, 2016. The population was characterized according to demographic data, species of fish associated with allergic reactions, and symptoms. The SPT wheal diameter and sIgE for fish and rGad c 1 were evaluated before acquisition of tolerance (T0) and afterwards (T1). RESULTS: The study population comprised 81 patients (68% male). Most reactions were triggered by hake (51%), mackerel (30%), and cod (26%). The most frequent manifestations were urticaria/angioedema (72%), gastrointestinal symptoms (35%), and eczema (33%); 42% of patients experienced anaphylaxis. At T0, the average sIgE values were as follows: cod, 32.2 kUA/L; sardine, 18.4 kUA/L; hake, 17.5 kUA/L; salmon, 13.9 kUA/L; tuna, 4.5 kUA/L; and rGad c 1, 22.9 kUA/L. In patients who acquired tolerance to at least 1 fish species (n=60; 74%), the mean value of rGad c 1 at T1 (5.1 kUA/L) was significantly lower than at T0 (16.8 kUA/L) (P=.001). Significant values were also recorded for the average diameter of the SPT wheal and the evaluations at T0 and T1 for hake (9.42 mm/3.79 mm) and salmon (7.8 mm/2.8 mm) (P=.002 and P=.026, respectively). CONCLUSION: The decrease in sIgE to rGad c 1 and the mean wheal diameter of SPT for hake and salmon can be used as markers of prognosis in the acquisition of tolerance by fish-allergic patients.

Peripheral Biomarkers of Inflammation in Depression: Evidence from Animal Models and Clinical Studies.

Depression is a highly prevalent psychiatric condition, with over 300 million sufferers, and is an important comorbidity for other conditions, like cardiovascular disorders or diabetes. Therapy is largely based on psychotherapy and/or pharmacological intervention, particularly aimed at altering neurotransmitter levels in the central nervous system, but inadequate response to treatment remains a significant clinical problem. Herein, evidence supporting a molecular link between inflammation and depression will be discussed, particularly the increased prevalence of depression in chronic inflammatory diseases and the evidence on the use of anti-inflammatory drugs to treat depression. Moreover, the potential for the levels of peripheral inflammatory molecules to act as depression biomarkers, in the diagnosis and monitoring of depression will be examined, considering clinical- and animal model-based evidence.

The Contribution of Inflammation to Autism Spectrum Disorders: Recent Clinical Evidence.

Autism comprises a complex and heterogeneous spectrum of neurodevelopmental disorders, usually termed autism spectrum disorders (ASD). It is more prevalent in males than females, and genetic and environmental factors are believed to account in similar percentages to the development of ASD. In recent years, the contribution of inflammation and inflammatory mediators to disease aetiology and perpetuation has been the object of intense research. In this chapter, inflammatory aspects that contribute to ASD are discussed, including abnormal microglia activation and polarization phenotypes, increased systemic levels of pro-inflammatory mediators, and altered patterns of immune cell response to activation stimuli. Also, inflammation in the context of gut microbiome and the impact of inflammation on gender prevalence of ASD are considered. Finally, treatment impact on inflammatory parameters and the potential for use of anti-inflammatory drugs, alone or in combination with antipsychotics, to manage ASD are examined.

Echocardiographic reference ranges for sedated healthy peccaries ( Tayassu tajacu, Linnaeus, 1758 ) / Valores de referência para catetos saudáveis sedados

Peccaries are wild mammals belonging to the Tayassu genus that are found almost everywhere in the Americas and have demonstrated great potential as an experimental model for scientific investigations. Twelve healthy adult animals were sedated to perform echocardiographic examinations in B, M and Doppler mode. The variables that exhibited statistically significant correlation coefficients with weight were LVFWd, LVIDd, LVIDs, E wave, A'RV, MAM, and TAPSE. The HR exhibited a negative relationship with the IVRT. The LA variable showed a positive correlation with the AO. The MAM exhibited correlations with the LVIDd and LVIDs. The TAPSE showed positive correlations with the E'RV and A'RV. The present study provides the first reference values for echocardiographic measurements in B, M and Doppler modes from peccaries anesthetized with ketamine and midazolam. Echocardiography was easy to perform in collared peccaries, and the collected data revealed values that can aid in their clinical management and conservation.(AU)

Catetos são mamíferos selvagens, pertencentes ao gênero Tayassu, encontrados em quase toda a América, os quais têm se destacado como modelos experimentais. Doze animais adultos saudáveis foram sedados para a realização de exames ecocardiográficos em modos B, M e Doppler. As variáveis que apresentaram coeficientes de correlação estatisticamente significativos em relação ao peso foram: LVFWd, LVIDd, LVIDs, onda E, A'RV, MAM, TAPSE. O HR mostrou correlação positiva com LA/AO, onda E, onda A, MAM, TAPSE e E/IVRT, e negativa com IVRT. Os valores de AVmax mostraram correlações positivas com onda E e onda A'. A variável LA mostrou correlação positiva com AO e correlação negativa com IVSd. A EF apresentou forte correlação com a relação de ondas FS, E/A, com A', E'RV. FS apresentou correlação positiva com a relação de onda E/A. O MAM mostrou correlação com LVIDd e LVIDs. TAPSE mostrou correlações positivas com E'RV e A'RV. O presente estudo forneceu os primeiros valores de referência para medições ecocardiográficas em modos B, M e Doppler de catetos anestesiados com ketamina e midazolam. O exame ecocardiográfico em catetos foi de fácil execução e os dados encontrados evidenciaram valores que podem auxiliar no seu manejo clínico e conservação.(AU)

In vivo and clinical application of strontium-enriched biomaterials for bone regeneration: A systematic review.

OBJECTIVES: This systematic review aimed to assess the in vivo and clinical effect of strontium (Sr)-enriched biomaterials in bone formation and/or remodelling. METHODS: A systematic search was performed in Pubmed, followed by a two-step selection process. We included in vivo original studies on Sr-containing biomaterials used for bone support or regeneration, comparing at least two groups that only differ in Sr addition in the experimental group. RESULTS: A total of 572 references were retrieved and 27 were included. Animal models were used in 26 articles, and one article described a human study. Osteoporotic models were included in 11 papers. All articles showed similar or increased effect of Sr in bone formation and/or regeneration, in both healthy and osteoporotic models. No study found a decreased effect. Adverse effects were assessed in 17 articles, 13 on local and four on systemic adverse effects. From these, only one reported a systemic impact from Sr addition. Data on gene and/or protein expression were available from seven studies. CONCLUSIONS: This review showed the safety and effectiveness of Sr-enriched biomaterials for stimulating bone formation and remodelling in animal models. The effect seems to increase over time and is impacted by the concentration used. However, included studies present a wide range of study methods. Future work should focus on consistent models and guidelines when developing a future clinical application of this element.Cite this article: N. Neves, D. Linhares, G. Costa, C. C. Ribeiro, M. A. Barbosa. In vivo and clinical application of strontium-enriched biomaterials for bone regeneration: A systematic review. Bone Joint Res 2017;6:366-375. DOI: 10.1302/2046-3758.66.BJR-2016-0311.R1.

Human Bone Marrow Mesenchymal Stem/Stromal Cells Preserve Their Immunomodulatory and Chemotactic Properties When Expanded in a Human Plasma Derived Xeno-Free Medium.

Due to their immunomodulatory and chemotactic properties, hMSC are being explored to treat immune-related diseases. For their use in human therapies, it is necessary to culture hMSC in xeno-free conditions. In this study, the impact that a xeno-free medium based on a human plasma derivate has on these properties was analysed. Bone marrow-derived hMSC preserved their immunosuppressive and immunostimulatory properties, as observed with in vitro assays with hMSC cocultured with mixed leukocyte reactions or with mitogen-stimulated leukocytes. Moreover, hMSC expanded in xeno-free medium were recruited by macrophages in both migration and invasion assays, which indicates that the cells maintained their chemotactic properties. These data suggest that xeno-free expanded hMSC preserved their immunomodulatory and chemotactic properties, indicating that the described xeno-free medium composition is a potential candidate to culture and expand hMSC for human cell therapies.

Pediatric eosinophilic esophagitis in Portugal.

SUMMARY: Eosinophilic esophagitis (EoE) is an increasingly frequent diagnosis in our clinical practice, mainly in pediatric age. Allergic responses to food and aeroallergens have been increasingly implicated in the etiology of this disease. We describe a retrospective data analysis of pediatric EoE patients followed in our Immunoallergology Department. Of the 25 children (22 male, average 10.8 years), 88% had prior history of rhinoconjunctivitis, 76% asthma, 48% eczema and 36% food allergy. After evaluation, we identified in 76% and 92% of patients food and aeroallergen sensitization, respectively; 68% had simultaneously food and inhalant sensitization and 96% had at least one positive test to aeroallergens or food allergens. The first (44%) and the most frequent (56%) symptom was dysphagia. The time between symptoms onset and the EoE diagnosis averaged 18.6 ± 29.4 months. A multidisciplinary approach is needed for a correct evaluation, intervention and follow-up of these patients.

Decellularized human colorectal cancer matrices polarize macrophages towards an anti-inflammatory phenotype promoting cancer cell invasion via CCL18.

Macrophages are frequently identified in solid tumors, playing important roles in cancer progression. Their remarkable plasticity makes them very sensitive to environmental factors, including the extracellular matrix (ECM). In the present work, we investigated the impact of human colorectal tumor matrices on macrophage polarization and on macrophage-mediated cancer cell invasion. Accordingly, we developed an innovative 3D-organotypic model, based on the decellularization of normal and tumor tissues derived from colorectal cancer patients' surgical resections. Extensive characterization of these scaffolds revealed that DNA and other cell constituents were efficiently removed, while native tissue characteristics, namely major ECM components, architecture and mechanical properties, were preserved. Notably, normal and tumor decellularized matrices distinctly promoted macrophage polarization, with macrophages in tumor matrices differentiating towards an anti-inflammatory M2-like phenotype (higher IL-10, TGF-ß and CCL18 and lower CCR7 and TNF expression). Matrigel invasion assays revealed that tumor ECM-educated macrophages efficiently stimulated cancer cell invasion through a mechanism involving CCL18. Notably, the high expression of this chemokine at the invasive front of human colorectal tumors correlated with advanced tumor staging. Our approach evidences that normal and tumor decellularized matrices constitute excellent scaffolds when trying to recreate complex microenvironments to understand basic mechanisms of disease or therapeutic resistance.

Octadecyl Chains Immobilized onto Hyaluronic Acid Coatings by Thiol-ene "Click Chemistry" Increase the Surface Antimicrobial Properties and Prevent Platelet Adhesion and Activation to Polyurethane.

Infection and thrombus formation are still the biggest challenges for the success of blood contact medical devices. This work aims the development of an antimicrobial and hemocompatible biomaterial coating through which selective binding of albumin (passivant protein) from the bloodstream is promoted and, thus, adsorption of other proteins responsible for bacterial adhesion and thrombus formation can be prevented. Polyurethane (PU) films were coated with hyaluronic acid, an antifouling agent, that was previously modified with thiol groups (HA-SH), using polydopamine as the binding agent. Octadecyl acrylate (C18) was used to attract albumin since it resembles the circulating free fatty acids and albumin is a fatty acid transporter. Thiol-ene "click chemistry" was explored for C18 immobilization on HA-SH through a covalent bond between the thiol groups from the HA and the alkene groups from the C18 chains. Surfaces were prepared with different C18 concentrations (0, 5, 10, and 20%) and successful immobilization was demonstrated by scanning electron microscopy (SEM), water contact angle determinations, X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR). The ability of surfaces to bind albumin selectively was determined by quartz crystal microbalance with dissipation (QCM-D). Albumin adsorption increased in response to the hydrophobic nature of the surfaces, which augmented with C18 saturation. HA-SH coating reduced albumin adsorption to PU. C18 immobilized onto HA-SH at 5% promoted selective binding of albumin, decreased Staphylococcus aureus adhesion and prevented platelet adhesion and activation to PU in the presence of human plasma. C18/HA-SH coating was established as an innovative and promising strategy to improve the antimicrobial properties and hemocompatibility of any blood contact medical device.

Three-dimensional scaffolds of fetal decellularized hearts exhibit enhanced potential to support cardiac cells in comparison to the adult.

A main challenge in cardiac tissue engineering is the limited data on microenvironmental cues that sustain survival, proliferation and functional proficiency of cardiac cells. The aim of our study was to evaluate the potential of fetal (E18) and adult myocardial extracellular matrix (ECM) to support cardiac cells. Acellular three-dimensional (3D) bioscaffolds were obtained by parallel decellularization of fetal- and adult-heart explants thereby ensuring reliable comparison. Acellular scaffolds retained main constituents of the cardiac ECM including distinctive biochemical and structural meshwork features of the native equivalents. In vitro, fetal and adult ECM-matrices supported 3D culture of heart-derived Sca-1(+) progenitors and of neonatal cardiomyocytes, which migrated toward the center of the scaffold and displayed elongated morphology and excellent viability. At the culture end-point, more Sca-1(+) cells and cardiomyocytes were found adhered and inside fetal bioscaffolds, compared to the adult. Higher repopulation yields of Sca-1(+) cells on fetal ECM relied on ß1-integrin independent mitogenic signals. Sca-1(+) cells on fetal bioscaffolds showed a gene expression profile that anticipates the synthesis of a permissive microenvironment for cardiomyogenesis. Our findings demonstrate the superior potential of the 3D fetal microenvironment to support and instruct cardiac cells. This knowledge should be integrated in the design of next-generation biomimetic materials for heart repair.

Ovarian response is a better predictor of clinical pregnancy rate following embryo transfer than is thin endometrium or presence of an endometrioma.

OBJECTIVES: To examine whether endometrial thickness and the presence of endometrioma are independent predictors of clinical pregnancy rate or simply associated with poor ovarian response (POR). METHODS: This was a retrospective cohort study assessing the first cycle of all women undergoing in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) in a university hospital in Brazil between January 2011 and December 2012. Only the first cycle of each woman within the study period was considered. Women over 40 years of age and those who used clomiphene citrate during controlled ovarian stimulation (COS) or did not undergo embryo transfer were excluded from analysis. POR was defined as ≤ three oocytes retrieved and a thin endometrium was defined as endometrial thickness ≤ 7.0 mm on the day of human chorionic gonadotropin (hCG) administration. We performed a multiple regression analysis to identify which of the following parameters were independent predictors of clinical pregnancy: age, number of oocytes retrieved, endometrial thickness or the presence of endometrioma. RESULTS: Within the study period, 787 women began COS, but 270 were excluded from analysis. Among the 517 women analyzed, those who achieved pregnancy were younger and yielded more oocytes. The proportion of POR was higher in women with a thin endometrium (17/57 (29.8%) vs 80/460 (17.4%); P = 0.03) and in women with endometrioma (15/39 (38.5%) vs 82/478 (17.2%); P = 0.002). The results of regression analysis showed that only age and the number of oocytes retrieved were independent predictors of pregnancy. Additionally, we observed higher clinical pregnancy rates in women with a thin endometrium from whom ≥ seven oocytes were retrieved (11/25 (44.0%)) compared to women with normal endometrial thickness (99/241 (41.1%)). Considering only women from whom ≥ four oocytes were retrieved, we observed reasonable pregnancy rates in those with a thin endometrium (14/40 (35.0%)) and in those with endometrioma (9/24 (37.5%)). CONCLUSION: Both a thin endometrium and the presence of endometrioma are associated with POR but are not important independent predictors of clinical pregnancy. Good pregnancy rates can be observed when these conditions are present in women with a good ovarian response.

Desempenho, consumo e morfometria in vivo de cordeiros Santa Inês alimentados com rações contendo torta de girassol em substituição ao farelo de algodão / Performance, consumption and in vivo morphometry of Santa Inês lambs fed diets containing sunflower cake in place of cottonseed meal

Este estudo foi conduzido com o objetivo de determinar o melhor teor de substituição da proteína do farelo de algodão pela proteína da torta de girassol em dietas para cordeiros Santa Inês, por meio da avaliação do consumo, ganho de peso, conversão alimentar e medidas morfométricas. Foram confinados, por 60 dias, 30 cordeiros da raça Santa Inês, machos inteiros, com idade e peso médio no início do experimento de 80 dias e 21,45±2,16kg, respectivamente. O delineamento experimental foi completamente casualizado, sendo os animais divididos em 5 tratamentos, com 6 repetições por tratamento, de acordo com a quantidade de proteína do farelo de algodão substituída pela proteína da torta de girassol (0, 20, 40, 60 e 80% de substituição). As variáveis peso final, consumo de matéria seca e conversão alimentar não foram afetadas pelas dietas experimentais, embora o ganho médio diário tenha sido inferior para as dietas com maior teor de torta de girassol. Para os parâmetros de consumo, verificou-se diferença apenas no consumo de extrato etéreo em relação ao peso metabólico (g/kg PV0,75), em que cordeiros recebendo dieta com maior teor de torta de girassol ingeriram maiores quantidades de extrato etéreo. Em relação às medidas morfométricas, houve decréscimo linear da altura de dorso e largura de garupa com a inclusão da torta de girassol na dieta. A substituição da proteína do farelo de algodão pela proteína da torta de girassol afetou negativamente o ganho de peso de cordeiros Santa Inês em confinamento. Porém, teve pouca ou nenhuma influência nos parâmetros de consumo e nas medidas morfométricas in vivo.(AU)

This study was conducted with the aim to determine the optimal level of protein replacement of cottonseed meal protein by sunflower cake in diets for Santa Inês lambs, through the evaluation of consumption, weight gain, feed conversion and morphometric measurements. Thirty Santa Inês lambs, bulls, with age and weight at the beginning of the experiment of 80 days and 21.45±2.16kg, respectively, were confined for 60 days. A completely randomized experimental design was used, where the animals were divided into 5 treatments with 6 replicates per treatment. For the treatments, the protein from the cottonseed meal was replaced by the protein of the sunflower cake (0, 20, 40, 60 and 80% of substitutions). For morphometric determinations, the following measurements were taken: pre-slaughter body length, leg length, leg perimeter, height of the dorsum; hip height; chest girth, hip width and chest width. Final weight, dry matter intake and feed conversion were not affected by the experimental diets, although the average daily gain was lower for diets with higher content of sunflower cake. For consumption parameters, there was a difference only in the consumption of ether extract in relation to metabolic weight (g/kg PV 0, 75), where lambs receiving diets with higher levels of sunflower cake ingested larger amounts of ether extract. Regarding the morphometric measurements, there was a linear decrease of the height of the dorsum and hip width with the inclusion of sunflower cake in the diet. The replacement of cottonseed meal protein by sunflower cake protein negatively affected the weight gain of Santa Inês lambs. However, it affected little or nothing the parameters of consumption and in vivo morphometric measurements.(AU)

Ultrasound guidance during embryo transfer: a systematic review and meta-analysis of randomized controlled trials.

OBJECTIVES: To summarize the current evidence on the effect of using ultrasound (US) guidance during embryo transfer (ET). METHODS: In this systematic review, we included randomized controlled trials examining the effect of the use of US guidance during ET; data from studies using the same catheter type in study arms were not pooled with the results from studies that used different catheter types. RESULTS: Twenty-one studies were included in the quantitative analysis: 18 compared 'US guidance' with 'clinical touch', of which one was subsequently excluded from the quantitative meta-analysis owing to a lack of available data, three studies compared transvaginal US guidance with transabdominal US guidance, and one study compared 'hysterosonometry before ET' with US guidance. Comparison of the use of US guidance with clinical touch, in studies that used the same catheter type in the study arms, indicated a benefit of using US guidance during ET on the rates of live birth (relative risk (RR), 1.48 (95% CI, 1.16-1.87)), based on two studies involving 888 women with moderate-quality evidence, and on the rates of clinical pregnancy (RR, 1.32 (95% CI, 1.18-1.46)), based on 13 studies involving 3641 women with high-quality evidence. However, when comparing the use of US guidance with clinical touch in studies that used different catheter types, the results suggest that using US guidance during ET has no effect on the rates of reproductive outcome: live birth (RR, 0.99 (95% CI, 0.83-1.19)), based on one study involving 1649 women with moderate-quality evidence; clinical pregnancy (RR, 1.04 (95% CI, 0.89-1.21)), based on five studies involving 2949 women with moderate-quality evidence. The estimates for the rate of miscarriage and for the other identified comparisons were imprecise. CONCLUSIONS: The available evidence suggests that there is a benefit of using US guidance during ET. However, both US-guided transfer and clinical touch should be considered acceptable, as the benefit of US is not large and should be balanced against the increased cost and need to change the catheter type. More studies are required before conclusions can be drawn regarding the effect of other techniques on reproductive outcome.

Desempenho de novilhos de corte em pastos de capim-tanzânia sob quatro alturas de desfolha / Performance of steers on pasture of Tanzania grass under four defoliation heights

Determinou-se a melhor altura de pastejo do capim-tanzânia (Panicum maximum Jacq. cv. Tanzânia) sob lotação contínua e com taxa de lotação variável na produção de bovinos de corte. A área experimental, de 12 hectares, foi dividida em piquetes de um hectare cada, com três repetições para cada altura - 20; 40; 60 e 80cm. Foram utilizados, em cada piquete, três bovinos machos, da raça Nelore, mais animais adicionais conforme necessidade de ajuste à altura pretendida. O delineamento experimental foi inteiramente ao acaso. A taxa de lotação apresentou comportamento linear negativo em razão das alturas de pastejo em todas as estações do ano, e o mesmo efeito foi observado com o ganho de peso vivo por hectare, enquanto o ganho médio diário apresentou comportamento contrário. Alturas de capim-tanzânia entre 40 e 60cm, sob lotação contínua, possibilitam bom ganho de peso médio diário em bovinos de corte e boa produção por hectare...

This study was carried out to evaluate the effects of four grazing heights under continuous stocking of Tanzânia grass (Panicum maximum Jacq. Cv. Tanzânia) on the stocking rate, performance and beef production per hectare. The experimental area was 12 hectares, divided into paddocks with one hectare each, with three replicates each (20, 40, 60 and 80 cm). There were three male Nellore bovines per paddock, adding animals as needed to adjust the desired heights in a completely randomized design. The stocking rate had a negative linear function of grazing heights in all seasons regarding yield per hectare, while the average daily gain showed opposite behavior. Apparently, Tanzânia grass heights between 40 and 60cm under continuous stocking enable good average daily weight gain in beef cattle, and high yield per hectare...