Advanced Maturation Research Articles

Quality of bananas harvested at different development stages and subjected to cold storage

ABSTRACT One of the biggest problems faced by banana growers is to determine the most appropriate harvest time. This study aimed to evaluate the post-harvest conservation of 'Prata-Anã' bananas harvested at 16, 17, 18, 19 and 20 weeks after the inflorescence emergence and subjected to 25 days of cold storage at the temperature of 13.5 ºC (± 1 ºC) and 90 % (± 5 %) of relative humidity. The bananas harvested at 19 and 20 weeks after the inflorescence emergence showed a more advanced maturity stage after 25 days of cold storage at 13.5 ºC. The bananas showed an increase in the content of soluble solids, as well as sugars, during the evaluations, after withdrawal from the cold room. Regardless of the fruit age, there was a decrease in the pH values over the 9 days of evaluation, after the 25-day storage period. The fruits at the ages of 19 and 20 weeks presented, on average, starch contents lower than those of the fruits with 16, 17 and 18 weeks. Fruits at the ages of 19 and 20 weeks showed higher extravasation levels on the second day of evaluation. Both the fruit harvest time and storage temperature directly influenced the post-harvest conservation and the physical and chemical characteristics of the 'Prata-Anã' bananas.

Advanced maturation of human cardiac tissue grown from pluripotent stem cells.

Cardiac tissues generated from human induced pluripotent stem (iPS) cells can serve as platforms for patient-specific studies of physiology and disease1–6. The predictive power of these models remains limited by their immature state1,2,5,6. We show that this fundamental limitation could be overcome if cardiac tissues are formed from early iPS-derived cardiomyocytes (iPS-CM), soon after the initiation of spontaneous contractions, and subjected to physical conditioning of an increasing intensity. After only 4 weeks of culture, these tissues displayed adult-like gene expression profiles, remarkably organized ultrastructure, physiologic sarcomere length (2.2 μm) and density of mitochondria (30%), the presence of transverse tubules (t-tubules), oxidative metabolism, positive force-frequency relationship, and functional calcium handling for all iPS cell lines studied. Electromechanical properties developed more slowly and did not achieve the stage of maturity seen in adult human myocardium. Tissue maturity was necessary for achieving physiologic responses to isoproterenol and recapitulating pathological hypertrophy, in support of the utility of this tissue model for studies of cardiac development and disease.

Morphological, anatomical and reproductive features of Loliolus (Loliolus) hardwickei (Gray, 1849) caught from the Vizhinjam Coast, Kerala, India

The morphological, anatomical and reproductive features of the little Indian squid, Loliolus (Loliolus) hardwickei (Gray, 1849) were studied based on samples collected from boat seine operated along the Vizhinjam Coast. The species was found to be a new entrant to the fishery along the Vizhinjam Coast. A total of 27 morphometric characters were measured and 13 indices were calculated and reported. Dorsal mantle length (DML) of the specimens ranged from 2.6 to 6.8 cm with a mean DML of 4.5±0.88 cm. The largest specimen observed was a male with DML 6.8 cm. Fully mature (Stage IV) specimens were found to have a higher gonadosomatic index (12.54±2.64 in females and 0.8±0.21 in males). Average length and width of spermatophores were 3.078±0.092 mm and 0.11±0.02 mm respectively. Samples with higher DML were in advanced maturity stages (Stage V). Size composition (DML) of samples revealed that 50% of squids were within the 4-5 cm size range.

Downstream effects of a large reservoir on the reproductive activity of Prochilodus hartii (Pisces: Prochilodontidae)

To analyze the effect of the Irapé reservoir on the reproduction of “curimba”, Prochilodus hartii, a migratory species, 503 specimens were collected during the reproductive period from February 2012 to January 2013. Specimens were collected from two river sections within the Jequitinhonha River Basin: Site 1 – a section of the Itacambiruçu River, a tributary of the Jequitinhonha River upstream of the Irapé reservoir; and Site 2 – a section of the Jequitinhonha River immediately downstream from the Irapé dam. Specimens (n = 270) were captured at Site 1 (160 males and 110 females), and 233 specimens at Site 2 (136 males and 97 females). The following were determined for each specimen: gonadosomatic index (GSI), hepatosomatic index (HSI), stomach repletion index (SRI), coelomic fat index (CFI) and condition factor (K). Both male and female fish were captured at all stages of gonadal maturation at Site 1, whereas only fish in the inactive and initial maturation stages were captured at Site 2. Length and mean GSI were greater (P < .05) at Site 1. Mean HSI and SRI of females was less in the advanced maturation stage. There were lesser values ​​for temperature and dissolved oxygen at Site 2 than 1. It is possible that the lesser values ​​for the physical and chemical characteristics of the water at Site 2 failed to promote advanced gonadal maturation, spawning, and spermiation of P. hartii, thus there are associations that are indicative of a negative effect of the Irapé reservoir on the reproduction of this migratory species.

Evolution of porosity and pore geometry in the Permian Whitehill Formation of South Africa – A FE-SEM image analysis study

Since the advent of technologically efficient exploitation of economic hydrocarbon reservoirs in shales, increasingly more research has been devoted to identifying and characterizing pore systems within shales. However, only a handful of these studies focused on the development of porosity in thermally mature unconventional reservoirs. In this study, the evolution of porosity and pore geometry in the Permian Whitehill Formation is addressed with the aid of ultrathin sections (2 × 3 cm, 10–20 μm thick) and field-emission scanning electron microscopy (FE-SEM) on samples with mean random vitrinite reflectance values ranging from 1.03 to 4.07 %Ro. We document a strong positive covariation of porosity and total organic carbon content (TOC) in all localities. However for samples with vitrinite reflectance values greater than 2.88 %Ro porosity per unit TOC decreased by over 25% relative to samples with lower thermal maturities. The positive covariation of thermal maturity and total porosity recorded here is unsurprising and have been documented previously in many gas shales. However, the dramatic decrease in porosity restricted to samples from localities that experienced advanced maturation (Ro > 2.88%) is viewed as an evidence that porosity decrease is directly related to late thermal decarboxylation of organic matter. This is supported by the presence of pores and micro-fractures infilled by fibrous grains, including carbonates, clays, silicates, and phosphates, and residual fluid inclusions. These grains were likely generated from re-precipitation of framework grains previously dissolved by organic acids (carboxylic, phenolic) that were generated during thermochemical decarboxylation of the OM. Our findings do not only fill important gaps in the understanding of organic pore development, including processes that create, preserve, and destroy porosity, the porosities described here are also key to gas transfer from shale matrix to induced fractures during fracture stimulation programs.

Preterm birth with placental evidence of malperfusion is associated with cardiovascular risk factors after pregnancy: a prospective cohort study.

Preterm birth (PTB) is associated with excess maternal cardiovascular disease risk. We considered that women with PTB and placental evidence of maternal malperfusion would be particularly affected. Pregnancy cohort study. Pittsburgh, PA, USA. Women with PTB (n = 115) and term births (n = 210) evaluated 4-12 years after pregnancy. Cardiometabolic risk markers were compared in women with prior PTB versus term births; pre-eclampsia and growth restriction cases were excluded. Placental evidence of maternal vascular malperfusion (vasculopathy, infarct, advanced villous maturation, perivillous fibrin, intervillous fibrin deposition), acute infection/inflammation (chorioamnionitis, funisitis, deciduitus) and villitis of unknown aetiology (chronic inflammation) was used to classify PTBs. Carotid artery intima-media thickness (IMT), fasting lipids, blood pressure (BP) and inflammatory markers measured after delivery. Women with PTB and malperfusion lesions had higher total cholesterol (+13.5 mg/dl) and systolic BP (+4.0 mmHg) at follow up compared with women with term births, accounting for age, race, pre-pregnancy BMI, and smoking (P < 0.05). Women with PTB and malperfusion accompanied by inflammatory lesions had the most atherogenic profile after pregnancy (cholesterol +18.7, apolipoprotein B + 12.7 mg/dl; all P < 0.05), adjusted for pre-pregnancy features. Carotid IMT was higher in this group (+0.037 cm, P = 0.031) accounting for pre-pregnancy factors; differences were attenuated after adjusting for BP and atherogenic lipids at follow up (+0.027, P = 0.095). PTBs with placental malperfusion were associated with an excess maternal cardiometabolic risk burden in the decade after pregnancy. The placenta may offer insight into subtypes of PTB related to maternal cardiovascular disease. Preterm births with placental malperfusion may mark women at higher cardiovascular disease risk.

Comparative study of sewage sludge co-composting with olive mill wastes or green residues: Process monitoring and agriculture value of the resulting composts

Abstract The viability of co-composting as a treatment approach for sewage sludge with organic wastes was investigated in the context of management aspects and the agricultural value of the final product. Two composting cycles (P1 and P2) were performed, in which sewage sludge (SS) was used to form two mixtures either with olive mill waste (OMW) or with green waste (GW), respectively. Co-composting of SS with both used organic wastes allowed obtaining hygienic compost with sufficient agronomic quality. Phenols accumulation from the mixture containing OMW caused an important decrease of pathogens within the compost. These products were characterized by a content of P and K that meet similar quality compared to commercial composts and showed an advanced maturation useable directly in agriculture. FTIR analyses and phytotoxicity tests were assessed to evaluate compost maturity. In fact, the germination index (GI) values for maize and tomato seeds, treated with P1, were 79.68% and 97.36%, respectively. However, they decreased to 74.45% and 81.45%, respectively using P2. Furthermore, compost application in peat amended at ratios equal to 30% and 50% improved the growth speed and fresh biomass of maize and tomato plants.

Induced Pluripotent Stem Cell (iPSC)-Based Mapping of Globin Expression throughout Human Erythropoietic Development

It has recently become clear that in vitro hematopoietic differentiation protocols using pluripotent stem cells mainly capture the primitive wave of hematopoietic development with limited induction of the definitive wave that gives rise to cells with adult-type characteristics such as the ability to express adult β-globin. Recent efforts from our group in optimizing our original differentiation protocol to derive more definitively patterned erythroid cells from iPSCs have resulted in a significant increase in β-globin transcripts as well as robust β-globin protein expression at the most mature stage of differentiation. To better quantify our progress in augmenting β-globin expression and to track globin ontogeny in real-time, we created a β-globin reporter iPSC line that allows for the mapping of β-globin expression throughout erythroid development.To create this tool, TALEN were used to target the β-globin locus in iPSCs where a promoterless GFP cassette was fused in frame to the first codon of the β-globin gene allowing for visualization of β-globin expression at single cell resolution via GFP expression.Interestingly, using our optimized protocol, only about 1% of cells exceeded the GFP detection threshold at the most mature stage of differentiation, suggesting that the several log-fold increase in β-globin transcripts seen at the population level as the cells progress through erythroid differentiation could be the result of high levels of β-globin transcription in just a small fraction of cells. Single cell RNA sequencing of GFP- and GFP+ sorted fractions showed significantly greater levels of β-globin transcripts in the GFP+ fraction in line with this hypothesis, thereby validating our reporter line as a tool to visualize and enrich for β-globin expressing cells.Despite lacking β-globin protein expression, the GFP- cells were not completely devoid of β-globin transcripts. As recent studies suggest that translation is dynamically controlled in maturing red blood cells, our results might indicate that posttranscriptional mechanisms could impact the translation of these globin transcripts. In fact, mining of the genes differentially expressed between both populations revealed several transcripts enriched in the GFP+ fraction that code for proteins involved in the dynamic translational control of transcripts essential for maturing red blood cells. These findings suggest advanced maturation of the GFP+ fraction, as well as a role for posttranscriptional mechanisms in the regulation of β-globin protein expression during erythroid development from iPSCs.Establishment of the developmental time frame of iPSC-derived erythroid cells is challenging as in vitro differentiation cultures lack the spatiotemporal separation of the different hematopoietic waves present in the embryo. Due to the possibility of multiple hematopoietic programs co-existing in one well, bulk expression analyses should be interpreted with caution when used to ascribe primitive or definitive characteristics to cell populations. Considering that the complexity of globin regulation in development might currently be underappreciated due to these factors, we also used single cell RNA sequencing to dissect the globin expression profiles of individual cells. Looking at the co-expression of different globins in individual cells, we found that the majority of the cells express a combination of embryonic (ε), fetal (γ) and adult (β) globins, indicative of a definitive yolk sac identity. Moreover, as β-globin transcripts increased, we observed a decrease in ε-globin transcripts, indicating that primitive/embryonic characteristics are gradually lost as cells gain more definitive/adult features.Further interrogation of the ‘β-globin expression signature’ that we distilled from the single cell RNA sequencing data will be instructive for future strategies aimed at increasing β-globin protein levels in iPSC-derived erythroid cells. Progress in this area will improve the resolution with which we can study hemoglobinopathies such as β-thalassemia and sickle cell disease and such strategies can then immediately be tested using the β-globin reporter iPSC line as a screening platform. DisclosuresNo relevant conflicts of interest to declare.

Effect of sourdough fermentation and baking process severity on bioactive fiber compounds in immature and ripe wheat flour bread

The dietary fiber complex of cereal whole grain is an important source of host beneficial molecules, such as: beta-glucans, fructans, resistant starch and arabinoxylans. The bioavailability of these compounds from whole grain products is generally dependent to different physiological and technological issues, likewise: genotype and maturation stage, dough preparation, fermentation options and baking process. This study regards the production of breads from cereal whole grains, taking into account two wheat genotypes harvested at mature and immature stages and processed through different conditions, varying temperature, time of baking and fermentation parameters. The aim is to highlight the different impact of these aspects on the amount and the bioavailability of the aforementioned four bioactive molecules. Our results showed that the best product, in terms of content of beneficial compounds, is that obtained from the flour of advanced maturation stage processed with brewer's yeast fermentation. Given the great interest in finding better technological processes to increase the daily dietary fiber intake, our findings could be very useful for the food industry to design the exact formulation and best process for the increase of bio-active molecules concentration in cereal whole grains products.

Role of FSH glycan structure in the regulation of Sertoli cell inhibin production.

Variations in follicle-stimulating hormone (FSH) carbohydrate composition and structure are associated with important structural and functional changes in Sertoli cells (SCs) during sexual maturation. The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. Recombinant human FSH (rhFSH) glycosylation variants were isolated according to their sialylation degree (AC and BA) and complexity of oligosaccharides (CO and HY). Native rhFSH stimulated inhibin α-subunit (Pro-αC) but did not show any effect on inhibin B (INHB) production in immature SCs isolated from 8-day-old rats. Activin A stimulated INHB and had a synergistic effect on FSH to stimulate Pro-αC. The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. Native rhFSH stimulated both Pro-αC and INHB in SCs at a more advanced maturation stage, isolated from 20-day-old rats. In these cells, all rhFSH glycosylation variants increased INHB and Pro-αC production, even in the presence of growth factors. The BA preparation exerted a more marked stimulatory effect on INHB and Pro-αC than the AC. Glycoforms bearing high mannose and hybrid-type oligosaccharides, HY, stimulated INHB and Pro-αC more effectively than those bearing complex oligosaccharides, CO, even in the presence of gonadal growth factors. These findings demonstrate the modulatory effect of FSH oligosaccharide structure on the regulation of inhibin production in the male gonad.

The effects of sGnRHa implants on Piaractus mesopotamicus female breeders. An approach addressed to aquaculture

In this study, we evaluated the potential use of sGnRHa slow-release implants (150 μg – Ovaplant®) for promoting an increased number of suitable Piaractus mesopotamicus females for induced spawning in a predictable period. To that, fshb, lhb, and gh gene expression, ovarian stereological evaluation and gonadal steroid concentrations were compared between treated and control females (n = 18) at 21, 35, and 115 days after sGnRHa implantation (DAI). The major differences between groups were observed at 35 DAI, when lhb and fshb were, respectively, up and downregulated for treated females compared to the controls, and the frequency of treated females in an advanced maturation class was higher (60%) than the controls (20%). Especially for promoting a significant change in the gonadotrophins gene expression levels, we demonstrated for the first time that sGnRHa slow-release implants has potential to be used to manipulate the reproductive cycle increasing the number of suitable Piaractus mesopotamicus females for induced spawning in a predictable period. However, a definitive protocol still depends on more studies involving a greater number of treatments and replicates.

Relative Age Effect and Academic Timing in American Junior College Baseball.

Previous research has shown that older athletes within age groupings are often perceived to be more talented simply due to advanced maturity, leading to biased selection in higher levels of sports competition, now commonly termed relative age effect (RAE). This study's goals were to determine whether (a) RAE influenced the selection of junior college baseball participants and (b) academic timing ( Glamser & Marciani, 1992 ), in which academic status determines age groupings more than strict age guidelines for college sports, influenced the formation of RAE. Participants were 150 junior college baseball players. Our results showed that RAE was only a significant factor, comparing the birth distribution of participants born before and after the midpoint of the participation year, when academic timing was also a factor in determining age groupings. In addition, the birth rate distribution, though not significantly different than expected, was greater only when those participants born during the expected participation year were included. The results of this study indicate that RAE could bear more influence among American student-athletes than was previously reported in that RAE in conjunction with academic timing does influence the selection of collegiate athletes.

The Effects of Maturity and Preservation Method on Nutrient Composition and Digestibility of Master Graze

The objective of experiment two was to evaluate the effects of maturity and harvest preservation method on Master Graze (MG) nutrient composition and digestibility. The MG was harvested on June 28 (Cut 1; 48 days), July 20 (Cut 2; 69 days), and August 12 (Cut 3; 92 days) and samples were then preserved using three different preservation methods (oven-dried hay (HAY), ensiled (SIL), or freeze dried (FD)) at each maturity. The collected samples were then analyzed for organic matter (OM), crude protein (CP), ether extract (EE), acid detergent fiber (ADF), neutral detergent fiber (NDF) and lignin. The OM content was similar for Cut 2 and Cut 3 and both were higher (P &lt; 0.01) than Cut 1 (92.4, 95.4 and 95.0% for cuts 1 to 3, respectively). The NDF content was highest (P &lt; 0.05) with Cut 3 (63.1%) and was least (P &lt; 0.05) with Cut 2 (60.1%). Cut 3 had a higher (P &lt; 0.01) lignin content compared to both Cut 1 and Cut 2 (3.5, 3.4 and 4.1%, respectively). Crude protein content was higher (P &lt; 0.01) for Cut 1 than Cut 2 and Cut 3 (8.0, 6.0, and 6.0%, respectively). The preservation method did not affect the CP or lignin content of the MG. The NDF content was highest (P &lt; 0.01) with HAY (65.8%) and was least (P &lt; 0.01) with FD (58.6%). The digestibility of the MG was measured under in situ conditions using one cow fitted with a rumen cannula. The ground oven-dried hay, ensiled and freeze dried samples were incubated in the rumen for 24 h using the mobile nylon bag techniques. The DM digestibility for Cut 1 and Cut 2 were higher (P &lt; 0.01) than Cut 3 (70.0, 67.0 and 60.3%, respectively). The NDF digestibility was highest for Cut 1 and decreased (P &lt; 0.01) with maturity (58.5, 51.6 and 43.8% for Cuts 1 to 3, respectively). Similarly, the CP digestibility was highest for Cut 1 and also decreased (P &lt; 0.01) with maturity (68.3, 56.1 and 46.5% for Cuts 1 to 3, respectively). The DM and OM digestibility was higher (P &lt; 0.04) with the FD in comparison to both HAY and SIL. Digestibility of NDF was not affected (P &gt; 0.05) by preservation methods, while the CP digestibility was higher (P &lt; 0.03) with the FD (61.6%) than the HAY (51.4%). In conclusion, the nutrient composition and digestibility of the MG were both affected by stage of the maturity and preservation method. The nutritional quality of MG decreased with advanced maturity and the MG digestibility was higher when FD preservation method was used.

Abstract 29: Chronic Stress Predisposes to Thrombosis by Abnormal Megakaryopiesis: Protective Effect of Apocynin

Introduction: Psychological stress (e.g. anxiety and depression) has been identified as an important trigger of acute coronary syndromes (ACS), as a consequence of enhanced coagulation and of hyper-reactive platelets. Changes in redox balance, alteration of genes regulating antioxidant systems, including NADPH oxidase, and increased production of reactive oxygen species (ROS) have been measured in both chronic stress and ACS. However, the mechanisms by which chronic stress affects platelet activation and predisposes to thrombosis are not well known. Hypothesis: We hypothesized that Apocynin, an inhibitor of NADPH oxidase influences the alteration of megakaryopoiesis and activation of platelets induced by chronic stress in mice. Methods and Results: We show the NADPH/NADP + ratio in bone marrow (BM) of mice exposed to forced swimming for 4 days (5 min twice/day) is markedly reduced compared to control mice, and that Apocynin treatment (2.4 mg/ml in drinking water for 4 days) prevents this alteration. Chronic stress leads to an abnormal megakaryopoiesis increasing the number of BM megakaryocytes (MKs) and affecting circulating platelets. MKs of stressed mice show an advanced maturation state (e.g. nuclear/cytoplasmic ratios and expression of CD42d), and an enhanced ability to produce ROS. Interestingly, a higher number of large and reticulated platelets with marked functional activation (e.g. integrin α IIb β3 and P-selectin expression, and platelet/leukocyte aggregates) is detected after chronic stress. In addition, Apocynin prevents ROS MKs generation and decreases the total number of MKs without affecting the percentage of CD42d + cells. Finally, the inhibitor of NADPH oxidase activity reduces the hyper-activation of platelets and the enhanced susceptibility to FeCl3-induced arterial thrombosis in stressed mice. Conclusion: Apocynin treatment, reducing ROS generation in MKs, restores the physiological bone marrow megakaryopoiesis and platelet behaviour, and it prevents the detrimental effect of chronic stress on atherothrombosis. These data suggest a potential use of NADPH oxidase inhibitors in the occurrence of thrombosis associated with chronic stress. Studies in human will verify the clinical impact of these findings.

Identification of a novel heterozygous mutation of the Aggrecan gene in a family with idiopathic short stature and multiple intervertebral disc herniation

Aggrecan is a critical proteoglycan component of the extracellular matrix of the growth plates and articular cartilage and has a key role in the biophysical and biomechanical properties of cartilage. Recently, heterozygous mutations in the ACAN gene, which encodes aggrecan, have been identified in patients with short stature and accelerated bone age. We herein report another family with a heterozygous ACAN mutation associated with idiopathic short stature along with accelerated bone age and early-onset herniation of the lumbar discs at the levels of L1/2 through L5/S1. Whole-exome sequencing identified a novel heterozygous frameshift mutation in the ACAN gene (c.1744delT; p.Phe582fs*69) in all of the affected family members but not in the unaffected one, providing further evidence that ACAN haploinsufficiency causes short stature with advanced bone maturation. In addition, we advocate early-onset multiple disc herniation as a novel phenotype associated with ACAN haploinsufficiency.

The Role of Inflammation in Cardiovascular Outcome

The aim of this review is to update the pathophysiological role of innate immune response in the cardiovascular (CV) disease outcomes, particularly focusing on coronary atherosclerosis and heart failure. Inflammatory processes comprised with the innate immunity reaction are believed to actively trigger CV disease development and final clinical events. For instance, by releasing proteases and neutrophil extracellular traps, neutrophil recruitment and activation might strongly influence atherosclerotic plaque stability. Similarly, neutrophils drive the early inflammatory response following a myocardial infarction. However, these cells contribute themselves to infarct healing by orchestrating monocyte/macrophage recruitment and polarization within the ischemic myocardium. Given their heterogeneity and plasticity, the balance between recruitment, proliferation, and polarization of monocyte/macrophage is a further leading determinant of advanced plaque maturation. Moreover, timely shift from a pro-inflammatory to a resolving macrophage phenotype may influence cardiac remodeling as well as development of heart failure (HF). Alongside macrophage recruitment and activation into the remote, non-ischemic myocardium also contributes to cardiac remodeling and HF development. Innate immune response is a tightly regulated process where a timely modulation of the balance between damaging and resolving properties critically impacts on CV outcome. Further progress may improve the determination of the prognostic relevance of inflammatory biomarkers on clinical CV outcome.

Defined Engineered Human Myocardium With Advanced Maturation for Applications in Heart Failure Modeling and Repair.

Advancing structural and functional maturation of stem cell-derived cardiomyocytes remains a key challenge for applications in disease modeling, drug screening, and heart repair. Here, we sought to advance cardiomyocyte maturation in engineered human myocardium (EHM) toward an adult phenotype under defined conditions. We systematically investigated cell composition, matrix, and media conditions to generate EHM from embryonic and induced pluripotent stem cell-derived cardiomyocytes and fibroblasts with organotypic functionality under serum-free conditions. We used morphological, functional, and transcriptome analyses to benchmark maturation of EHM. EHM demonstrated important structural and functional properties of postnatal myocardium, including: (1) rod-shaped cardiomyocytes with M bands assembled as a functional syncytium; (2) systolic twitch forces at a similar level as observed in bona fide postnatal myocardium; (3) a positive force-frequency response; (4) inotropic responses to β-adrenergic stimulation mediated via canonical β1- and β2-adrenoceptor signaling pathways; and (5) evidence for advanced molecular maturation by transcriptome profiling. EHM responded to chronic catecholamine toxicity with contractile dysfunction, cardiomyocyte hypertrophy, cardiomyocyte death, and N-terminal pro B-type natriuretic peptide release; all are classical hallmarks of heart failure. In addition, we demonstrate the scalability of EHM according to anticipated clinical demands for cardiac repair. We provide proof-of-concept for a universally applicable technology for the engineering of macroscale human myocardium for disease modeling and heart repair from embryonic and induced pluripotent stem cell-derived cardiomyocytes under defined, serum-free conditions.

Opportunistic spawning of tropical anguillid eels Anguilla bicolor bicolor and A. bengalensis bengalensis

Information on the spawning migration, spawning ecology and life history of tropical eels in the Indo-Pacific region is very limited. The physiological and morphological characteristics of tropical freshwater eels, Anguilla bicolor bicolor and A. bengalensis bengalensis collected in Malaysia were examined in relation to their downstream migration patterns. A total of 455 eels were collected over monthly intervals between February 2014 and January 2016 and we examined both gonadosomatic index and gonad histology features. In both species, close positive relationships between advanced maturation stages and eye, fin, gonadosomatic indexes were found in males and females. A. bengalensis bengalensis was found to be larger and heavier than A. bicolor bicolor at the time of seaward migration. The final stage of maturation for seaward spawning migration occurred throughout the year in A. bicolor bicolor, although that of A. bengalensis bengalensis was estimated to six months due to the limited number of samples. These results suggest that year-round spawning in the open ocean occurs in the tropical eel. This non-seasonal spawning ecology is notably different from that of temperate eels, which are known to follow a well-defined spawning season, with spawning migrations generally taking place during autumn months.

Neonatal outcomes following preterm birth classified according to placental features

Preterm birth has staggering health implications, and yet the causes of most cases are still unknown. Placental features have been understudied as an etiology for preterm birth, and the association between placental pathologic lesions and neonatal outcomes are incompletely understood. We sought to characterize births according to placental pathology and relate these to adverse neonatal outcomes. We studied 20,091 births (15,710 term and 4381 preterm) with placental evaluations. Births were classified according to the presence or absence of placental lesions consistent with malperfusion (vasculopathy, infarct, advanced villous maturation, perivillous fibrin, fibrin deposition) and intrauterine inflammation/infection (chorioamnionitis, funisitis, vasculitis). Outcomes were gestational week of delivery, birthweight z-score, neonatal respiratory distress syndrome, and intraventricular hemorrhage. Among all preterm births, evidence of placental malperfusion was identified more often than inflammation/infection (50.6% vs 27.3%, P< .0001). Placental malperfusion was associated with reduced fetal growth (adjusted birthweight z-score, -0.83, P < .0001) and lesions of inflammation/infection were associated with earlier delivery (adjusted difference -2.08 weeks, P < .0001) than those with no lesions. When both placental lesions were present, earlier delivery (adjusted difference -2.28 weeks, P < .0001) and reduced fetal growth (adjusted birthweight z-score difference, -0.24, P= .001) were observed more often than when neither lesion was present. Findings were similar when restricted to cases of spontaneous preterm birth. Intraventricular hemorrhage was higher in preterm births with malperfusion lesions than cases with no lesions (7.6% vs 3.4%; odds ratio, 1.98; confidence interval, 1.18-3.32), accounting for gestational age and other covariates. Placental pathology provides important insight into subtypes of preterm birth with adverse neonatal outcomes. Co-occurrence of malperfusion and inflammation/infection, especially among spontaneous preterm births, may be a novel pattern of placental injury linked to severe adverse outcomes.

Relationship between Heifer Carcass Maturity and Beef Palatability

ObjectivesOur objective was to determine the relationship between carcass maturity and beef palatability of strip loin and outside round steaks.Materials and MethodsLeft sides of A (n = 30), B (n = 30), and C (n = 30) maturity heifer carcasses under 30 mo of age by dentition were used. Average skeletal maturities of the groups were A67, B49, and C48. Carcasses were selected to ensure similar marbling scores across maturity groups, and average marbling scores were Sm94, Mt02, and Mt01 for A, B, and C maturity carcasses, respectively. Beef strip loins (IMPS 180) and outside (bottom) rounds (IMPS 171B) obtained from these carcasses were purchased from AB Foods (Toppenish, WA) and transported to the University of Idaho Meat Science Laboratory. Following a 14-d aging period, wholesale cuts were removed from vacuum packaging and ischiatic heads were removed from outside rounds to produce trimmed flats. Two 2.54 cm-thick steaks were cut from the trimmed flats and anterior ends of strip loins. Steaks were used to measure Warner-Bratzler shear force (WBSF), cook loss, insoluble and total collagen, and consumer sensory attributes. Steaks used for WBSF were weighed, cooked on open hearth broilers to an internal temperature of 40°C, then turned and cooked to a final internal temperature of 71°C. Cooked steaks were re-weighed to determine cook loss and cooled to 4°C. Six cores (1.27-cm diameter) were then removed from each steak parallel with the muscle fibers and then sheared perpendicular to muscle fiber orientation using a Warner-Bratzler shear machine (GR Manufacturing, Manhattan, KS). Samples from cooked steaks were frozen at –20°C and used to determine insoluble and total collagen. Sensory panel steaks were frozen at –20°C after aging, and allowed to thaw at 4°C for 24 h prior to the consumer panel. Steaks were cooked as previously described and four 1.27-cm × 1.27-cm × steak thickness cubes were obtained from each steak. Separate sensory panels were conducted for strip loin and outside round steaks. Consumers (n = 72 panelists per panel) evaluated cooked samples for overall acceptability, tenderness, juiciness, and flavor using a 9-point scale (9 = like extremely, 1 = dislike extremely). Using an incomplete block design, panelists evaluated 5 samples from the 3 maturity groups. Data were analyzed using the Mixed Model procedure of the Statistical Analysis System (SAS Inst. Inc., Cary, NC). Significance was determined at P < 0.05 and data were considered trending at P < 0.10.ResultsHeifer carcass maturity did not significantly affect WBSF or cook loss for either outside round or strip loin steaks (P > 0.23). Likewise, insoluble and total collagen were not different for either steak type from A, B, or C maturity carcasses (P > 0.89). Strip loin steaks from C maturity carcasses tended to have higher overall acceptability (P = 0.08) and juiciness (P = 0.09) than steaks from B maturity carcasses, but steaks from B and C maturity carcasses did not differ from strip loin steaks obtained from A maturity carcasses (P > 0.11.). No differences in tenderness or flavor were observed due to maturity (P > 0.24). Similarly, maturity had no effect on sensory characteristics of outside round steaks (P > 0.30).ConclusionIn conclusion, advanced skeletal maturity does not decrease palatability of carcasses from cattle under 30 mo of age.