Improved maintenance of adult rat hepatocytes in a new serum-free medium in the presence or absence of barbiturates

Summary For serum-free primary culture of adult rat hepatocytes, a synthetic medium DM-160 and rat-tail collagen were selected for the basal medium and for the culture substratum, respectively. Barbiturates, such as phenobarbital and 1-ethyl-5-isobutylbarbiturate, efficiently supported survival of hepatocytes and maintained their morphologic features at lower concentrations under the serum-free conditions than under the serum-supplemented conditions. However, the hepatocyte survival rates under the serum-free conditions were lower than those under the serum-supplemented conditions in the presence or absence of barbiturates. Supplementation of the basal medium with a combination of five groups of factors (5Fs), such as eight amino acids (Ala, Arg, Gly, Ile, Met, Phe, Pro, and Trp), two unsaturated fatty acids (linoleate and oleate), a protease inhibitor (aprotinin), three vitamins (A, C, and E), and five trace elements (Mn, Fe, Cu, Zn, and Se), improved the hepatocyte survival under the serum-free conditions in the presence or absence of barbiturates. In other words, the serum could be completely substituted by the 5Fs. Hepatocyte cultures maintained in the 5Fs-suppelemented basal medium showed excellent induction of tyrosine aminotransferase activity in response to dexamethasone in the presence or absence of barbiturates. The efficiency of the 5Fs-supplemented basal medium for maintaining hepatocytes was not inferior to those of other media in common use with hepatocytes, such as Williams' medium E and Waymouth's medium MB-752/1. In conclusion, maintenance of functional hepatocytes in serum-free primary culture could be improved by use of the new medium preparation (the 5Fs-supplemented DM-160) in the presence of barbiturates. This work was supported by a grant no. 61771923 from the Ministry of Education, Science and Culture of Japan.     

Biological studies including population growth parameters of the predatory miteAmblyseius barkeri [Acarina.: Phytoseiidae] at 25°C in the laboratory

Amblyseius barkeri (Hughes) fed onThrips tabaci (Lind.) at 25°C showed an average duration of 2.2, 0.8 and 3.2 days for the egg, larval and nymphal stages, with mortalities at 1.0, 1.0 and 3.1%, respectively. Females represented 63% of the population and required multiple matings for optimal fertility. The oviposition period was 20.3 days and the average oviposition rate 2.3 eggs per day. The intrinsic rate of increase was 0.22 per day. The expected life span was 29.6 days for ♀♀ and 27.4 days for ♂♂.A. barkeri ♂♂ and ♀♀ both consumed 3.3 nymphs of thrips per day (mean value for the feeding stages). The larva does not take up food. In the absence of thripsA. barkeri was able to consume two-spotted spider mites,Tetranychus urticae (Koch), and their eggs, adult broad mites,Polyphagotarsonemus latus (Banks), and pollen of various plants. Cannibalism was observed when food was lacking. Certain morphological features, egglaying, mating and predatory behaviour are described.      

Bionomics ofTrioxys indicus [Hym.: Aphidiidae], a parasitoid ofAphis craccivora [Hem.: Aphididae]: 31. Effect of host haemolymph on the numerical response of the parasitoid

The influence of kairomones on the numerical response of the parasitoidTrioxys indicus against its hostAphis craccivora at its varying density was studied. The kairomones (applied as aqueous extract of the host) significantly enhanced the rate of parasitisation and multiplication and the area of discovery of the parasitoid and also the K-values of mortality of the host at all parasitoid densities introduced (1, 2, 4, 8, 12 and 16 parasitoids) into troughs having about 200 hosts. The sex-ratio of F₁ offspring decreased at lower parasitoid densities and remained more or less unchanged at higher parasitoid densities after the application of kairomones. The present findings indicate that if kairomones are applied properly, the number of hosts destroyed by a stimulated parasitoid will be about 200, twice the number reported earlier, thus fewer parasitoids will be needed to regulate an estimated population of the hosts.      

Bioluminescence in oceanology

For analytical purposes bioluminescence can be used in three main ways: 1. luminescence measurement of bioluminescent system components isolated in vitro; 2. determination of luminous organisms' reaction to the in vivo test-action; 3. measurement of bioluminescence in marine ecological systems. The majority of the reports of this Symposium are dealing with the first two topics. The aim of our presentation is to draw attention to the third one. The possibilities of bioluminescent analysis are wider than its traditional scheme of applications in the laboratory, when the emitting system is withdrawn from a native source and is placed in a cuvette of the light measuring device. The reverse scheme is also possible, i.e. the device can be introduced into light emitting system such as a marine biocenosis–the community of the sea inhabitants–where we obtain a highly sensitive and rapid means of gaining the information on the vital activity of marine ecosystems, i.e. their spatial structure, rhythms, man's influence upon them, etc. The present communication will consider the possibilities of this form of bioluminescent analysis.     

关于茭白黑粉菌的正名问题

茭白黑粉菌在文献中出现Yenia esculenta(P.Henn.)Liou及Ustilago esculenta P.Henn.两个名字。现经形态发生学的研究以明确新名字是否有效。此菌冬孢子没有休眠期,孢子成熟后在适宜的环境中即可萌发。萌发的最适pH值为6;最适温度为25％;不需要外界光的诱导,在黑暗条件下也能正常萌发;培养基中的营养成份对孢子萌发的速率有一定影响。冬孢子在不同的环境条件下,其萌发形态及方式都是稳定的:孢子萌发时产生有隔的担子或先菌丝(promycelium)。该先菌丝起初可以是无隔的,但生长到一定程度,便产生分隔,分隔多为2—4个。初生的先菌丝和小孢子是单核的,但成熟的先菌丝、小孢子及短菌丝细胞内均是双核的。可以认为,该菌的双核期在整个生活史中占有相当长的时间,它只有短暂的单核期。没有发现先菌丝、小孢子彼此之间或相互间融合的现象;未见到双核的融合及减数分裂过程。茭白黑粉菌在人工组合培养基上生成厚壁的孢子,较自然界中的冬孢子为大,但萌发方式则相同。最后作者等认为茭白黑粉菌仍应保留在Ustilago属中,而Yenia esculenta(P.Henn.)Liou的名字是无效的。     

小麦颖果果皮绿色层内同化物的合成与分配

小麦幼嫩颖果中,果皮内侧发育出由内表皮与亚表皮组成的一薄层绿色组织。显微与亚显微结构观察表明,虽绿色层只接受到自然光照的1/4～1/8,叶绿体仍能正常发育,叶绿素含量与叶绿体数量均高出旗叶。大量胞间连丝联接相邻的绿色细胞,并在一定时期形成开放的胞间通道,显然有利于同化物的快速胞间运输。离体颖果饲喂~14CO_2试验证明,新合成的同化物从绿色细胞输向胚珠。绿色层产生的同化物可能通过合点端的珠心进入胚乳或通过珠孔直接汇聚到胚珠和分化中的原胚。     

新疆黑粉菌的初步研究

作者于1986年参加巴尔鲁克山真菌考察,并到阿尔泰山地、伊犁谷地、吐鲁番等地采得黑粉菌标本70余份,鉴定为8属36种,13种为我国新纪录,它们是石竹叶苔炭黑粉(Anthracoidea caryophylleae Latour.),卡里炭黑粉(Anthracoidea karii(Liro) Nannf.),范基灰黑粉(Anthracoidea vankyiNannfeldt),短柄草腥黑粉(Tilletia olida (Riess) Schr(?)t.),剪股颍条黑粉(Urocystis agrostidis (Lavrov.) Zundel),黑麦草条黑粉(Urocystis bo-livari Bubák & Fragoso),雀麦条黑粉(Urocystis bromi (Lavrov) Zundel),拂子茅条黑粉(Urocystis calamagrostidis (Lavrov) Zundel),茅香条黑粉(Urocystis hierochloae (Murashk) Vánky),尼氏条黑粉(Urocystis nevodo-vskyi Schwarzman),早熟禾条黑粉(Urocystis poae(Liro)Padw.& Khan),报春花条黑粉(Urocystis primulicola P Magn.),和泡斑黑粉(Ustilagopustulata (D.C.) Winter     

人胃癌不同周期细胞膜表面ConA受体的分布与侧向运动

本文研究了人胃低分化粘液性腺癌细胞MGC 80-3不同周期时相中ConA受体的分布与侧向运动。MGc 80-3细胞经同步化培养,用F-ConA标记。被标记细胞中G_1、S和G_2期呈不连续的分布,但它们之间又存在显著的差异。M期呈较均匀的强荧光分布(与其它时相细胞比较)。荧光漂白恢复方法测定ConA受体复合物侧向运动表明:各个周期时相之间不仅运动方式不同,而且运动速率也有显著差异。M期与G_1期主要表现出扩散型运动;而S期与G_2期表现为流动型运动。G_1期的扩散系数大干M期的;S期的流动速率大于G_2期的。但可动分子百分比以G_2期最高。这些结果表明了ConA受体的动力学性质。它受到细胞周期的调节。     

黑腹果蝇P转座因子研究 Ⅰ.我国黑腹果蝇的P-M测定及其地理分布

采用性腺败育(GD不育)作为标准检定方法。对我国20个地方的黑腹果蝇的P因子活性和细胞型进行了测定。结果表明我国北部沿海城市为Q型;南部沿海和内地皆为M型。各地的M品系所产生的GD不育能力各不相同,但表现出与地理位置相关的梯度变化。这一变化规律为研究我国黑腹果蝇的P因子起源及P和M品系的形成提供了重要的理论依据。