Opiate binding sites and endogenous opioids in Bufo viridis oocytes

Binding sites with high affinity for [3H]naloxone, but not for [3H]morphine and [3H] (D-Ala2, D-Leu5) enkephalin, have been found in membranes of Bufo viridis oocytes. The binding is reversible and saturable. Bound [3H]naloxone is easily displaced both by unlabeled naloxone and bremazocine, much worse by morphine and SKF 10,047; (D-Ala2, D-Leu5) enkephalin and beta-endorphin practically fail to displace [3H]naloxone. Scatchard analysis is consistent with the existence of two classes of binding sites with Kd 15 nM and 10(3) nM. The number of binding sites with high affinity for naloxone is 16 pmol/mg protein of homogenized oocytes which is 20-50-fold higher than in, toad or rat brain. Oocyte extract displaces [3H]naloxone bound with oocytes' membranes and inhibits electrically evoked contractions of the rabbit vas deferens. This inhibition is reversed by naloxone. It is suggested that compounds similar to opiate kappa-agonists exist in oocytes. It cannot be ruled out that they participate via specific receptors in the regulation of oocyte maturation and egg development.     

血管平滑肌细胞的分化与血管内壁的构建

血管平滑肌细胞(vascular smooth muscle cells,VSMCs)的发育与血管壁的构建是目前相关领域中的重要学科前沿．国内外同行的工作多集中在血管发育初始阶段内皮细胞及其前体细胞在血管新生中的作用、调节因素及生物学机制．VSMCs参与血管壁早期构建,特别是VSMCs的募集与分化机制已经成为血管新生研究中的一个新领域． 本期发表的《 抑制Rac1蛋白活化阻碍胚胎发育早期血管新生 》(见696～701页)报道了韩雅玲教授及其合作者在这一领域取得的最新研究结果．Rac1是真核细胞内重要的一类信号传递分子,在细胞信号传递过程中发挥分子开关作用．他们采用胚胎干细胞(ESCs)为模型,建立稳定表达持续型Rac1和显性失活型Rac1编码序列的小鼠ESCs并制备胚胎小体,诱导分化后观察其对内皮细胞分化和迁移的影响,发现抑制Rac1可以干扰血管内皮细胞连接成血管网状结构,细胞骨架F-actin排列紊乱,细胞的迁移受到明显抑制,表明Rac1在胚胎早期血管发育过程中与内皮细胞的迁移有关[1]． 近年来,韩雅玲教授及其研究集体在VSMCs发育与血管构建、胚胎干细胞来源的拟胚体血管平滑肌发育与血管新生机制以及胚胎主动脉VSMCs起源等方面开展了研究,取得了一系列有价值的成果[2～11],可能为闭塞性和增生性血管病的发生及防治提供理论依据和候选基因．详见“相关链接”．     

Systematic butchering of fallow deer (Dama) at the early middle Pleistocene Acheulian site of Gesher Benot Ya'aqov (Israel)

Three assemblages of fallow deer (Dama sp.) bones excavated from the early middle Pleistocene (oxygen isotope stage 18) layers of the Acheulian site of Gesher Benot Ya'aqov, Israel, furnish evidence of systematic and repeated exploitation of complete carcasses by hominins. The excellent state of preservation of the bones and the presence of only minimal signs of carnivore involvement permit an investigation of the role of hominins as the primary agents responsible for the damage to these bones. Hominin expertise in dealing with fallow deer carcasses is manifested by cut marks, percussion marks, and hack marks on the bones. The archaeozoological analysis of the anatomical position and frequency of these marks suggests that carcass processing followed systematic practices that reflect an in-depth knowledge of fallow deer anatomy and a consistent behavioral strategy. These assemblages represent one of the earliest examples of methodological butchering practices in Eurasia. The evidence of carcass processing observed at Gesher Benot Ya'aqov resembles that seen in late Pleistocene sites in Israel, which were inhabited by modern humans. We interpret the Gesher Benot Ya'aqov data as indicating that the Acheulian hunters at the site (1) were proficient communicators and learners and (2) possessed anatomical knowledge, considerable manual skill, impressive technological abilities, and foresight.     

A new type of anvil in the Acheulian of Gesher Benot Ya‘aqov,Israel

We report here on the identification and characterization of thin basalt anvils, a newly discovered component of the Acheulian lithic inventory of Gesher Benot Ya‘aqov (GBY). These tools are an addition to the array of percussive tools (percussors, pitted stones and anvils) made of basalt, flint and limestone. The thin anvils were selected from particularly compact, horizontally fissured zones of basalt flows. This type of fissuring produces a natural geometry of thick and thin slabs. Hominins at GBY had multiple acquisition strategies, including the selection of thick slabs for the production of giant cores and cobbles for percussors. The selection of thin slabs was carried out according to yet another independent and targeted plan. The thinness of the anvils dictated a particular range of functions. The use of the anvils is well documented on their surfaces and edges. Two main types of damage are identified: those resulting from activities carried out on the surfaces of the anvils and those resulting from unintentional forceful blows (accidents de travaille). Percussive activities that may have been associated with the thin anvils include nut cracking and the processing of meat and bones, as well as plants.     

The Early-Middle Pleistocene faunal assemblages of Gesher Benot Ya'aqov: Inter-site variability

This study investigates faunal remains from the site of Gesher Benot Ya‘aqov, analyzing how hominins may have utilized vertebrates and exploring paleoenvironments during the Early and Middle Pleistocene. Based on analyses of a range of vertebrates, results show that the species distribution of terrestrial vertebrates (microvertebrates and mammals) at the site of Gesher Benot Ya‘aqov shows relative stasis above the Matuyama-Brunhes Boundary (MBB) (0.78 Ma). However, consistency in faunal remains at the site reflecting stasis does not seem to reflect accurately the paleoenvironment. Marked changes are demonstrable in the lake-margin sedimentary background, archaeological remains, and agents of accumulation and damage, as well as in the density of medium-large mammals. This study emphasizes the significance of studying assemblages as a whole rather than the species representation on its own.     

Culture and cognition in the Acheulian industry: a case study from Gesher Benot Ya'aqov

The Acheulian presence in the Dead Sea Rift and its environs is characterized by the discontinuity of its cultural manifestations. Nevertheless, the long stratigraphic sequences of the Acheulian Technocomplex provide a unique opportunity for synergetic examination along a temporal trajectory. Hominin cognitive and cultural behaviour are studied at Gesher Benot Ya'aqov through analyses of lithic, palaeontological and palaeobotanical assemblages, as well as the Early-Middle Pleistocene environment, ecology and climate. The study attempts to reconstruct reduction sequences of some major artefact groups at the site, which include raw material acquisition, production, technology, typology, usage and discard. Experimental archaeology illustrates artefact mobility on the palaeo-landscape. Strategies of biomass-exploitation are studied in detail, with other aspects yielding additional information on hominin subsistence and adaptive responses to their environment. The cultural marker of fire and the spatial association of selected categories of finds are integrated in the general synthesis, allowing reconstruction of the cultural and cognitive realm of Acheulian hominins. The synthesis attempts to reassess the abilities, social structure, subsistence and adaptability to the changing environment of hominins in the Levantine Corridor.     

Binding Adaptation of GS-441524 Diversifies Macro Domains and Downregulates SARS-CoV-2 de-MARylation Capacity

Viral infection in cells triggers a cascade of molecular defense mechanisms to maintain host-cell homoeostasis. One of these mechanisms is ADP-ribosylation, a fundamental post-translational modification (PTM) characterized by the addition of ADP-ribose (ADPr) on substrates. Poly(ADP-ribose) polymerases (PARPs) are implicated in this process and they perform ADP-ribosylation on host and pathogen proteins. Some viral families contain structural motifs that can reverse this PTM. These motifs known as macro domains (MDs) are evolutionarily conserved protein domains found in all kingdoms of life. They are divided in different classes with the viral belonging to Macro-D-type class because of their properties to recognize and revert the ADP-ribosylation. Viral MDs are potential pharmaceutical targets, capable to counteract host immune response. Sequence and structural homology between viral and human MDs are an impediment for the development of new active compounds against their function. Remdesivir, is a drug administrated in viral infections inhibiting viral replication through RNA-dependent RNA polymerase (RdRp). Herein, GS-441524, the active metabolite of the remdesivir, is tested as a hydrolase inhibitor for several viral MDs and for its binding to human homologs found in PARPs. This study presents biochemical and biophysical studies, which indicate that GS-441524 selectively modifies SARS-CoV-2 MD de-MARylation activity, while it does not interact with hPARP14 MD2 and hPARP15 MD2. The structural investigation of MD?GS-441524 complexes, using solution NMR and X-ray crystallography, discloses the impact of certain amino acids in ADPr binding cavity suggesting that F360 and its adjacent residues tune the selective binding of the inhibitor to SARS-CoV-2 MD.