Improved blastocyst development of single cow OPU-derived presumptive zygotes by group culture with agarose-embedded helper embryos

<p>Abstract</p> <p>Background</p> <p>The <it>in vitro </it>culture of presumed zygotes derived from single cow ovum pick-up (OPU) is important for the production of quality blastocysts maintaining pedigree. The aim of the present study was to evaluate the agar chip-embedded helper embryo coculture system for single cow OPU-derived zygotes by assessing embryo quality.</p> <p>Methods</p> <p>Cumulus oocyte complexes (COCs) were collected from Hanwoo cows with high genetic merit twice a week using the ultra-sound guided OPU technique and from slaughterhouse ovaries. The Hanwoo cow COCs and slaughterhouse ovaries were matured <it>in vitro</it>, fertilized <it>in vitro </it>with thawed Hanwoo sperm and cultured for 24 h. The presumed zygotes were subsequently placed in three different culture systems: (1) control OPU (controlOPU) with single cow OPU-derived presumed zygotes (2~8); (2) agar chip-embedded slaughterhouse helper embryo coculture (agarOPU) with ten presumed zygotes including all presumed zygotes from a cow (2~8) and the rest from agar chip-embedded slaughterhouse presumed zygotes (8~2); and (3) slaughterhouse <it>in vitro </it>embryo production (sIVP) with ten slaughterhouse ovary-derived presumed zygotes, each in 50 μL droplets. Day 8 blastocysts were assayed for apoptosis and gene expression using real time PCR.</p> <p>Results</p> <p>The coculture system promoted higher blastocyst development in OPU zygotes compared to control OPU zygotes cultured alone (35.2 vs. 13.9%; P < 0.01). Genes predicted to be involved in implantation failure and/or embryo resorption were down-regulated (P < 0.05) in control OPU zygotes (<it>CD9</it>, 0.4-fold; <it>AKRAB</it>1, 0.3-fold) and in cocultured zygotes (<it>CD9</it>, 0.3-fold; <it>AKRAB</it>1, 0.3-fold) compared to sIVP blastocysts (1.0-fold). Moreover, genes involved in implantation and/or normal calf delivery were up-regulated (P < 0.05 to P < 0.01) in control OPU zygotes (<it>PGSH</it>2, 5.0-fold; <it>TXN</it>, 4.3-fold; <it>PLAU</it>, 1.7-fold) and cocultured zygotes (<it>PGSH</it>2, 14.5-fold; <it>TXN</it>, 3.2-fold; <it>PLAU</it>, 6.8-fold) compared to sIVP (1.0-fold) blastocysts. However, the expression of <it>PLAC8, TGF-β1, ODC1</it>, <it>ATP5A1 </it>and <it>CASP3 </it>did not differ between the three culture groups.</p> <p>Conclusions</p> <p>Results show that the agar chip-embedded helper embryo coculture system enhances developmental competence and embryo quality in cultures of limited numbers of high pedigree single cow OPU presumed zygotes.</p

Barrier Tissue Macrophages: Functional Adaptation to Environmental Challenges

Macrophages are found throughout the body, where they have crucial roles in tissue development, homeostasis and remodeling, as well as being sentinels of the innate immune system that can contribute to protective immunity and inflammation. Barrier tissues, such as the intestine, lung, skin and liver, are exposed constantly to the outside world, which places special demands on resident cell populations such as macrophages. Here we review the mounting evidence that although macrophages in different barrier tissues may be derived from distinct progenitors, their highly specific properties are shaped by the local environment, which allows them to adapt precisely to the needs of their anatomical niche. We discuss the properties of macrophages in steady-state barrier tissues, outline the factors that shape their differentiation and behavior and describe how macrophages change during protective immunity and inflammation

Simple clinical criteria to identify sepsis or pneumonia in neonates in the community needing treatment or referral.

BACKGROUND: Sepsis, meningitis and pneumonia annually kill 1.1 million neonates in developing countries; most deaths occur at home. OBJECTIVES: To develop simple clinical criteria, enabling health workers in communities to identify neonates with potentially fatal sepsis; and to identify the danger signs alerting mothers to seek care. METHODS: In a field trial in 39 villages in Gadchiroli, India, trained health workers visited all neonates at home 8 times during the first 28 days of life, recording signs and outcome without interventions during 1995-1996 and with home-based management of sick neonates during 1996-1999. An independent neonatologist assigned the cause of death. We use the term "sepsis" to include sepsis, meningitis and pneumonia. We evaluated 31 signs as predictors of 43 sepsis deaths among 3567 neonates. We also evaluated mothers' observations as the danger signs to seek care. RESULTS: Simultaneous presence of any 2 of 7 signs (reduced or stopped sucking; weak or no cry; limbs becoming limp; vomiting or abdominal distension; baby cold to touch; severe chest indrawing; umbilical infection) predicted sepsis death with sensitivity 100%, specificity 92%, positive predictive value 27.2% and negative predictive value 100% in the nonintervention period. The criteria identified 10.6% of the neonates in the community as suspected sepsis, at a mean of 5.4 days before death. The criteria remained valid in the postintervention period. Any 1 of the 5 maternally observed danger signs (reduced sucking, drowsy or unconscious, baby cold to touch, fast breathing and chest indrawing) gave 100% sensitivity and identified 23.9% neonates for seeking care. CONCLUSION: These criteria identify neonates in the community who are at risk for dying of infection with excellent sensitivity, specificity and negative predictive value but a moderate positive predictive value. They can be used by health workers to select sick neonates for treatment or referral. One potentially fatal case would be treated per 4 presumptive cases treated

Neurocomputational mechanisms of confidence in self and others

Estimating confidence in the decision making ability of others is important for cooperative behaviour. Here the authors combine computational modelling and fMRI to investigate how the brain supports this process

Confidence matching in group decision-making

Most important decisions in our society are made by groups, from cabinets and commissions to boards and juries. When disagreement arises, opinions expressed with higher confidence tend to carry more weight1,2 . Although an individual’s degree of confidence often reflects the probability that their opinion is correct3,4, it can also vary with task-irrelevant psychological, social, cultural and demographic factors5–9. Therefore, to combine their opinions optimally, group members must adapt to each other’s individual biases and express their confidence according to a common metric10–12. However, solving this communication problem is computationally difficult. Here we show that pairs of individuals making group decisions meet this challenge by using a heuristic strategy that we call ‘confidence matching’: they match their communicated confidence so that certainty and uncertainty is stated in approximately equal measure by each party. Combining the behavioural data with computational modelling, we show that this strategy is effective when group members have similar levels of expertise, and that it is robust when group members have no insight into their relative levels of expertise. Confidence matching is, however, sub-optimal and can cause miscommunication about who is more likely to be correct. This herding behaviour is one reason why groups can fail to make good decisions10–12