ON and OFF retinal ganglion cells differentially regulate serotonergic and GABAergic activity in the dorsal raphe nucleus

The dorsal raphe nucleus (DRN), the major source of serotonergic input to the forebrain, receives excitatory input from the retina that can modulate serotonin levels and depressive-like behavior. In the Mongolian gerbil, retinal ganglion cells (RGCs) with alpha-like morphological and Y-like physiological properties innervate the DRN with ON DRN-projecting RGCs out numbering OFF DRN-projecting RGCs. The DRN neurons targeted by ON and OFF RGCs are unknown. To explore retino-raphe anatomical organization, retinal afferents labeled with Cholera toxin B were examined for association with the postsynaptic protein PSD-95. Synaptic associations between retinal afferents and DRN serotonergic and GABAergic neurons were observed. To explore retino-raphe functional organization, light-evoked c-fos expression was examined. Light significantly increased the number of DRN serotonergic and GABAergic cells expressing c-Fos. When ON RGCs were rendered silent while enhancing the firing rate of OFF RGCs, c-Fos expression was greatly increased in DRN serotonergic neurons suggesting that OFF DRN-projecting RGCs predominately activate serotonergic neurons whereas ON DRN-projecting RGCs mainly target GABAergic neurons. Direct glutamatergic retinal input to DRN 5-HT neurons contributes to the complex excitatory drive regulating these cells. Light, via the retinoraphe pathway can modify DRN 5-HT neuron activity which may play a role in modulating affective behavior

Safety and tolerability of a single dose T0001 in Chinese healthy adult volunteers: a first-in-human ascending dose study

T0001 is the first mutant of etanercept with a higher affinity to tumor necrosis factor α (TNF-α) than etanercept. In order to investigate the safety and tolerability of T0001, a study was carried out in healthy Chinese subjects. A first-in-human, dose escalation study was conducted in healthy Chinese subjects. Fifty-six subjects were divided into six dose cohorts (10 mg, 20 mg, 35 mg, 50 mg, 65 mg and 75 mg) to receive a single subcutaneous injection of T0001. Safety and tolerability assessment were based on the records of vital signs, physical examinations, clinical laboratory tests, 12-lead electrocardiograms and adverse events (AEs). All subjects were in good compliance and none withdraw due to AEs. No serious AEs occurred. A total of twenty-three AEs in sixteen subjects were recorded, and eighteen of these AEs were believed to be related to T0001. The most frequently reported AEs were injection site reactions and white blood cell count increase. All these AEs were of mild to moderate intensity and most of them recovered spontaneously within 14 days. In this study, no dose-limiting toxicity was observed, and the maximum tolerated dose was identified as 75 mg. T0001 was considered safe and generally well tolerated at doses up to 75 mg in healthy Chinese volunteers

Вихретоковый анизотропный термоэлектрический первичный преобразователь лучистого потока

Представлена оригинальная конструкция первичного преобразователя лучистого потока, который может служить основой для создания приемника неселективного излучения с повышенной чувствительностью

Maternal control of early mammalian embryogenesis

This thesis explores and discusses biochemical changes during mammalian oogenesis with the aim of providing novel information, at the transcriptomic and proteomic levels, about events and processes that help to support, direct and protect the genomic integrity of the early mammalian embryo. PIWIs are crucial guardians of genome integrity, particularly in germ cells. While mammalian PIWIs have been primarily studied in mouse and rat, a homologue for the human PIWIL3 gene is absent in the Muridae family, and hence the unique function of PIWIL3 in germ cells cannot be effectively modeled by mouse knockouts. Herein, in chapter 2, we investigated the expression, distribution and interaction of PIWIL3 in bovine oocytes. We localized PIWIL3 to mitochondria, and demonstrated that PIWIL3 expression is stringently controlled both spatially and temporally before and after fertilization. Moreover, we identified PIWIL3 in a mitochondrial-recruited three-membered complex with TDRKH and PNLDC1, and demonstrated by mutagenesis that PIWIL3 N-terminal arginine modifications are required for complex assembly. Finally, we sequenced the piRNAs bound to PIWIL3-TDRKH-PNLDC1 and report here that about 50% of these piRNAs map to transposable elements, recapitulating the important role of PIWIL3 in maintaining genome integrity in mammalian oocytes. In chapter 2, microinjection was used for the investigation of PIWIL3 localization and function. While siRNA introduction by microinjection can be useful to study the function of maternally expressed genes in oocytes, injection is a rather invasive procedure in that it damages the plasma membrane and disturbs the cell organelle distribution. Therefore, in chapter 3 we examined the effect of introducing non-specific small RNAs into oocytes by microinjection, at the single cell transcriptome level. Injection of non-specific siRNA resulted in differential expression of 119 transcripts, of which 76 were down-regulated. Gene ontology analysis revealed that the differentially regulated genes were enriched in the biological processes of ATP synthesis, molecular transport and regulation of protein polyubiquitination. This study establishes a background effect of the microinjection procedure that should be borne in mind by those using microinjection to manipulate gene expression in oocytes. Based on the results of chapter 3, a group of mRNAs with functions related to plasma membrane interaction are significantly altered by microinjection, which raises the question of the importance of plasma membrane proteins during oocyte maturation and fertilization. In chapter 4, we used an optimized plasma membrane protein purification method followed by phospho enrichment and mass spectrometry to investigate the oolemma phospho(proteome) in cattle GV and MII stage oocytes and zygotes. We identified a number of protein complexes that connect with the plasma membrane, including the Arp2/3, ERM and SCMC complexes. Moreover, we detected multiple new phospho-sites that were not previously reported. In addition, we found that PALM3, PB41L2 and TACC3 increased significantly at both the protein and phospho-protein levels during oocyte maturation. Our research validated an effective method for plasma membrane protein identification from samples of limited protein content, and also provided general knowledge on plasma membrane proteins activated during oocyte maturation and fertilization

Maternal control of early mammalian embryogenesis

This thesis explores and discusses biochemical changes during mammalian oogenesis with the aim of providing novel information, at the transcriptomic and proteomic levels, about events and processes that help to support, direct and protect the genomic integrity of the early mammalian embryo. PIWIs are crucial guardians of genome integrity, particularly in germ cells. While mammalian PIWIs have been primarily studied in mouse and rat, a homologue for the human PIWIL3 gene is absent in the Muridae family, and hence the unique function of PIWIL3 in germ cells cannot be effectively modeled by mouse knockouts. Herein, in chapter 2, we investigated the expression, distribution and interaction of PIWIL3 in bovine oocytes. We localized PIWIL3 to mitochondria, and demonstrated that PIWIL3 expression is stringently controlled both spatially and temporally before and after fertilization. Moreover, we identified PIWIL3 in a mitochondrial-recruited three-membered complex with TDRKH and PNLDC1, and demonstrated by mutagenesis that PIWIL3 N-terminal arginine modifications are required for complex assembly. Finally, we sequenced the piRNAs bound to PIWIL3-TDRKH-PNLDC1 and report here that about 50% of these piRNAs map to transposable elements, recapitulating the important role of PIWIL3 in maintaining genome integrity in mammalian oocytes. In chapter 2, microinjection was used for the investigation of PIWIL3 localization and function. While siRNA introduction by microinjection can be useful to study the function of maternally expressed genes in oocytes, injection is a rather invasive procedure in that it damages the plasma membrane and disturbs the cell organelle distribution. Therefore, in chapter 3 we examined the effect of introducing non-specific small RNAs into oocytes by microinjection, at the single cell transcriptome level. Injection of non-specific siRNA resulted in differential expression of 119 transcripts, of which 76 were down-regulated. Gene ontology analysis revealed that the differentially regulated genes were enriched in the biological processes of ATP synthesis, molecular transport and regulation of protein polyubiquitination. This study establishes a background effect of the microinjection procedure that should be borne in mind by those using microinjection to manipulate gene expression in oocytes. Based on the results of chapter 3, a group of mRNAs with functions related to plasma membrane interaction are significantly altered by microinjection, which raises the question of the importance of plasma membrane proteins during oocyte maturation and fertilization. In chapter 4, we used an optimized plasma membrane protein purification method followed by phospho enrichment and mass spectrometry to investigate the oolemma phospho(proteome) in cattle GV and MII stage oocytes and zygotes. We identified a number of protein complexes that connect with the plasma membrane, including the Arp2/3, ERM and SCMC complexes. Moreover, we detected multiple new phospho-sites that were not previously reported. In addition, we found that PALM3, PB41L2 and TACC3 increased significantly at both the protein and phospho-protein levels during oocyte maturation. Our research validated an effective method for plasma membrane protein identification from samples of limited protein content, and also provided general knowledge on plasma membrane proteins activated during oocyte maturation and fertilization

PIWIL3 Forms a Complex with TDRKH in Mammalian Oocytes

P-element induced wimpy testis (PIWIs) are crucial guardians of genome integrity, particularly in germ cells. While mammalian PIWIs have been primarily studied in mouse and rat, a homologue for the human PIWIL3 gene is absent in the Muridae family, and hence the unique function of PIWIL3 in germ cells cannot be effectively modeled by mouse knockouts. Herein, we investigated the expression, distribution, and interaction of PIWIL3 in bovine oocytes. We localized PIWIL3 to mitochondria, and demonstrated that PIWIL3 expression is stringently controlled both spatially and temporally before and after fertilization. Moreover, we identified PIWIL3 in a mitochondrial-recruited three-membered complex with Tudor and KH domain-containing protein (TDRKH) and poly(A)-specific ribonuclease-like domain containing 1 (PNLDC1), and demonstrated by mutagenesis that PIWIL3 N-terminal arginines are required for complex assembly. Finally, we sequenced the piRNAs bound to PIWIL3-TDRKH-PNLDC1 and report here that about 50% of these piRNAs map to transposable elements, recapitulating the important role of PIWIL3 in maintaining genome integrity in mammalian oocytes

Multiagent learning of coordination in loosely coupled multiagent systems

Multiagent learning (MAL) is a promising technique for agents to learn efficient coordinated behaviors in multiagent systems (MASs). In MAL, concurrent multiple distributed learning processes can make the learning environment nonstationary for each individual learner. Developing an efficient learning approach to coordinate agents\u27 behaviors in this dynamic environment is a difficult problem, especially when agents do not know the domain structure and have only local observability of the environment. In this paper, a coordinated MAL approach is proposed to enable agents to learn efficient coordinated behaviors by exploiting agent independence in loosely coupled MASs. The main feature of the proposed approach is to explicitly quantify and dynamically adapt agent independence during learning so that agents can make a trade-off between a single-agent learning process and a coordinated learning process for an efficient decision making. The proposed approach is employed to solve two-robot navigation problems in different scales of domains. Experimental results show that agents using the proposed approach can learn to act in concert or independently in different areas of the environment, which results in great computational savings and near optimal performance

Emotional multiagent reinforcement learning in spatial social dilemmas

Social dilemmas have attracted extensive interest in the research of multiagent systems in order to study the emergence of cooperative behaviors among selfish agents. Understanding how agents can achieve cooperation in social dilemmas through learning from local experience is a critical problem that has motivated researchers for decades. This paper investigates the possibility of exploiting emotions in agent learning in order to facilitate the emergence of cooperation in social dilemmas. In particular, the spatial version of social dilemmas is considered to study the impact of local interactions on the emergence of cooperation in the whole system. A doublelayered emotional multiagent reinforcement learning framework is proposed to endow agents with internal cognitive and emotional capabilities that can drive these agents to learn cooperative behaviors. Experimental results reveal that various network topologies and agent heterogeneities have significant impacts on agent learning behaviors in the proposed framework, and under certain circumstances, high levels of cooperation can be achieved among the agents