In vitro mammalian mutagenesis as a model for genetic lesions in human cancer

Recently in vitro assays of mutagenesis have been criticized as being poorly predictive of long-term in vivo rodent assays of carcinogenicity. Questions have also been raised concerning the relevance of rodent assays to human risk. In vitro assays using mammalian cells can detect most types of genetic lesions thought to be important in human malignant disease. Molecular and cytogenetic analyses of mutations induced by a variety of genotoxic compounds at the heterozygous thymidine kinase locus in mouse lymphoma cells indicate that this in vitro assay does indeed register the range of genetic lesions recently found in a wide variety of human tumors. The types and complexity of the induced lesions are reflected in mutant colony phenotype in a compound-specific fashion. These studies point to the use of appropriate in vitro mammalian mutagenesis assays as new model systems for dissecting the genetic lesions important in human carcinogenesis, and as a means of determining the potential for compounds to induce such lesions.     

猕猴桃属(Actinidia)植物的离体种质保存

本文研究了猕猴桃属植物离体种质资源保存的方法。在一年中于盛夏时采取的植物材料在离体培养时能得到最好的效果;改进的剥离茎尖的方法使污染率大大降低,在MS附加BA 0.5,Z 0.1,GA 0.1～0.5 mg/L,蔗糖3％,琼脂0.55％的培养基中茎尖生长良好并且不产生不定芽。通过茎尖培养方法已保存了10多个种,40多个猕猴桃的离体种质。     

工程抗体的体外成熟

工程抗体在诊断或治疗方面有其优点,但制备的各种工程抗体的亲和力通常较原亲本抗体的低,为方便在体外迅速地改善工程抗体的生物特性,目前已发展了一系列针对抗体库构建,筛选和阳性克隆鉴定等与抗体外成熟密切相关的方法和技术。     

In vitro effects of Celiptium and MR 14504 on mature rat Leydig cell testosterone production

Percoll-purified mature rat Leydig cells have been used to evaluate the testicular toxicity of two highly potent intercalating agents (Celiptium and MR 14505). Testosterone secretion in the absence and in the presence of human chorionic gonadotropin (hCG) was measured to assess Leydig cell function. Celiptium and MR 14504 induce time- and dose-related inhibitory effects on the production of testosterone by Leydig cells, both in the presence and in the absence of hCG, whatever the concentration of hCG used. We have observed that MR 14504 is about 5 times more potent as an inhibitor of rat Leydig cell steroidogenesis than Celiptium without inducing any cell toxicity. The present study indicates that the Leydig cell is an additional potential site for the primary toxic effects of these drugs in the adult rat testis.     

In vitro translation of the genome of a small RNA virus from Trichoplusia ni

The genomic RNA of a member of the “Nudaurelia β virus” group functioned as a mRNA in vitro. The translation products included a protein, which comigrated with the single virus capsid protein, and a stable 100 × 10³ MW protein, which was synthesized by cleavage of a precursor protein. No precursor proteins were involved in synthesis of the putative capsid protein. Attempts to inhibit proteolytic cleavage did not result in the appearance of a product corresponding to the entire coding capacity of the genome.     

In vitro flowering of orchids

Abstract

Flowering is the most elusive and fascinating of all plant developmental processes. The ability to induce flowering in vitro in orchids would reduce the relatively long juvenile phase and provide deeper insight into the physiological, genetic and molecular aspects of flowering. This review synthesizes all available studies that have been conducted on in vitro flowering of orchids with the objective of providing valuable clues as to the mechanism(s) that is possibly taking place.     

In vitro fertilization of pig oocytes matured in vitro

The development of in vitro fertilization (IVF) techniques in pigs as well as in other species is of great importance because of the possible applications of this technology in different research fields. Methods of IVF vary in different incubation periods and temperatures, in the hormone concentrations used, and in the treatment of the sperm samples. It has been particularly difficult to succeed in the achievement of fertilization in the pig. In the present study we used FSH and LH concentrations of 2 IU/ml for oocyte maturation, an incubation temperature of 37°C, and dilution of spermatozoa for capacitation, and we achieved a high fertilization rate (50 to 75%) with no cases of polyspermy.     

番木瓜的离体繁殖

建立番木瓜离体繁殖体系.用0.1%HgCl2溶液对番木瓜的新生嫩茎进行消毒,适宜的消毒时间为12min,1mm茎尖的成苗率达到87.6%.随蔗糖浓度的提高,番木瓜试管苗的株高显著降低,增殖系数显著增加.在附加IBA0.3mg/L的1/2MS培养基上新梢的生根率达到89.3%,试管苗大规模移栽的成活率达90%以上.基因型显著地影响番木瓜离体增殖的效率.     

Developmental capacity of rat embryos produced by in vivo or in vitro fertilization

This work compares the ability of rat zygotes fertilized in vitro or in vivo to develop into viable embryos. All oocytes were from adult cyclic females. After the first cleavage, the zygotes were transferred to oviducts of pseudopregnant recipients. Their fate was examined on day 13 at laparotomy and again on day 20. Ninety-five of 146 in vivo fertilized zygotes developed into normal sized 13-day fetuses and 72 (55%) to apparently normal near-term fetuses. Forty-six of 135 in vitro fertilized zygotes developed up to day 13, and 30 (24%) developed to term. It appears that the probability that in vitro fertilized rat zygotes will develop into viable embryos is about half the chance of in vivo fertilized zygotes. Since the two types of zygotes were morphologically identical, the morphological appearance of the two-cell stage is not an adequate criterion for judging developmental potential.     

In vitro flowering in cassava (Manihot esculenta Crantz)

Meristem-derived plantlets of cassava (Manihot esculenta Crantz) were induced to flower in vitro. Five genotypes out of 13 consistently responded to our culture conditions giving rise to male or female flowers. Male flowers contained anthers in which meiosis occurred and apparently normal pollen grains were formed.     

大花卷丹的组织培养及限制生长保存

离体保存是植物种质保存的重要手段之一,为实现对大花卷丹的保护性利用,本文对其组织培养体系及限制生长离体保存技术进行了研究。结果表明,在常温（23±2）℃、光照强度约为40μmol.m-2.s-1、光照时间14h.d-1的条件下,大花卷丹鳞片在MS＋6-BA1.0mg.L-1＋NAA0.2mg.L-1培养基中生长情况较好,能直接诱导芽,且小鳞茎的生长速度较快。将诱导出的小鳞茎切割后,接种到1/2MS＋NAA0.5mg.L-1＋活性炭2g.L-1生根培养基2～3周即能生根,生长状况良好。提高MS培养基中蔗糖达90和110g.L-1时可以抑制其生长,能够保存大花卷丹试管苗10个月,保存过程中生长正常,株高生长缓慢,但根长势较快。在蔗糖浓度90g.L-1基础上再添加30g.L-1甘露醇的培养基能进一步抑制试管苗根的生长。6个月后,转移到正常培养基上培养均能恢复生长,其鳞片在诱导培养基上能正常分化。因此,采用大花卷丹鳞片组织培养可以形成种苗,在培养基中添加高蔗糖浓度和甘露醇可以使其试管苗保存1年以上。     

In vitro study of elements in herbal remedies

Decreased glucose tolerance is a first sign of diabetes mellitus and therefore rigorous control must be taken in carbohydrate and lipid metabolisms. Herbal remedies (lyophilized extracts of Myrtilli folium and Phaseoli fructus sine seminibus (L1), Myrtilli folium, Phaseoli fructus sine seminibus, and Salviae folium (L2) are traditionally used in mid-European folk medicine and in common adjuvant therapy for the prevention of complications in type 2 diabetes. Significant iron (355.7±13.8 mg/kg) and zinc (84.73±1.83 mg/kg) concentration was found in L1 and chromium (3.82±2.71 mg/kg) in L2. Ion concentrations in teas made from L1 and L2 are relatively low because the quantities of metal ions in teas do not cover the daily need, although the teas are good sources for some elements. According to the Recommended Daily Allowances, the tea of L1 is a good source for iron and manganese, whereas for chromium, the tea of L2 is better. For evaluating the element bioavailability, an in vitro dialysis system was applied to determine the element transfer from tea of the lyophilized sample to the plasma (buffer pH=7.4). Measurements showed that the elements transferred between 6.90% (iron from tea of L2) and 90.05% (chromium from tea of L2) through the membrane from teas to the plasma. Metal ions in teas of herbal remedies might contribute to the favorable therapeutic effect of preventing complications, because they might transfer through the membranes in relatively high percentages.     

蒙自凤仙花的离体保存

对蒙自凤仙花茎尖的离体保存研究结果表明：培养温度在12℃、18℃和24℃,每天12h光照(2000 lx),添加1％～5％甘露醇的MS培养基对茎尖的生长有抑制作用。最适保存条件为12℃,甘露醇含量为3％,保存1年后存活率为100％;存活的幼苗及茎尖接种于增殖培养基,仍能正常生长、增殖和生根。     

Isolation and visualization of Met-72-positive, metastatic variants present in B16 melanoma tumor masses

Metastatic variants of the B16 melanoma displaying high experimental metastatic potential have been shown to express high levels of a 72,000-dalton glycoprotein (Met-72) on their cell surface (Kimura AK, Xiang J: J Nat Can Inst 76:1247-1253, 1986). Monoclonal antibodies (MoAb) directed against the Met-72 determinant have been used in this study as immunohistochemical reagents on preparations of fresh B16 melanoma tumors and their metastases. These immunohistochemical analyses have utilized frozen sections, impression smears, and cytospin preparations of fresh tumors harvested at various time points during tumor growth, to view the presence and location of Met-72-positive metastatic variants within tumor masses. Biotinylated anti-Met-72 MoAbs were reacted with freshly dissociated tumor cells from a B16 melanoma ovarian metastasis. These cells were then reacted with fluorescein isothiocyanate (FITC)-streptavidin and analyzed by flow cytometry. A discrete population of positively staining cells was detected and isolated by cell sorting techniques. Met-72-positive cells were then cloned and reanalyzed after several weeks of in vitro expansion and found to have high experimental metastatic potential to ovaries. Frozen sections of subcutaneous tumors and their metastases were analyzed by immunoperoxidase techniques. A consistent finding in these studies has been that the few tumor cells which showed high intensity of Met-72 staining were positioned perivascularly and at the invading front of B16 melanoma tumors.     

桂花茎尖离体培养体系的建立

以桂花(Osmanthus fragrans Lour.)的休眠芽?萌动芽和嫩枝的茎尖为外殖体,进行离体培养研究,筛选其最适培养基。结果表明:3种外殖体的适宜初代培养基不同,其中:休眠芽为B5 7.0 mg.L-16-BA 0.05 mg.L-1NAA 3%蔗糖;萌动芽为B5 5.0 mg.L-16-BA 0.05 mg.L-1NAA 3%蔗糖;嫩枝为B5 3.0 mg.L-16-BA 0.05 mg.L-1NAA 3%蔗糖;但适宜的继代培养基(B5 2.0 mg.L-16-BA 0.05 mg.L-1NAA 3%蔗糖)和生根培养基(B5 2.0 mg.L-1NAA 3%蔗糖)相同。     

低氧大鼠脑线粒体体外转录活性的研究

目的：探讨低氧对大鼠脑线粒体DNA表达的影响及其与能量生成的关系。方法：雄性Wistar大鼠随机分为3组：急性低氧组（AH）、慢性低氧组（CH）和对照组,其中急、慢性低氧组动物分别连续暴露于模拟海拔4000m高原3d(AH)和40d（CH）。分离脑线粒体,分别测定线粒体体外转录活性、F0F1－ATP酶活性以及ATP对线粒体体外转录的影响。结果：急性低氧大鼠脑线粒体体外转录活性及F0F1－ATP酶活性显著降低,慢性低氧时有所回升,两者呈线性相关。ATP对大鼠脑线粒体体外转录活性呈双相效应。结论：低氧时脑线粒体转录活性改变可能参与低氧抑制线粒体能量代谢的机制,ATP可能通过反馈作用对线粒体转录进行微调。     

In vitro culture of ovaries of a viviparous gall midge

Summary Ovaries of the viviparous pedogenetic gall midgeHeteropeza pygmaea can be cultured in hemolymph obtained from X-ray-sterilized larvae of the same species. In this culture medium, formation of follicles is essentially the same as in vivo, and sometimes female larvae develop from these follicles. The ovaries of such larvae, in their turn, have been cultured in vitro to produce larvae. In this way, in vitro development from oogonium to larva has been maintained for several generations. When using hemolymph obtained from larvae grown under different conditions, the in vitro cultured ovaries produce a second type of egg which probably is male-determined. Ovarian development in vitro has been studied with differential interference contrast optics and time-lapse cinemicrography. This work was supported by the Swiss National Science Foundation Grant No. 3.2010.73.     

In vitro bone formation on coral granules

Summary We investigated the ability of fetal rat bone cells isolated after collagenase digestion to differentiate in vitro and to produce a mineralized matrix on coral granules. Scanning electron microscopy examination of the surface of the seeded coral granules revealed that cells attached, spread, and proliferated on the material surface. Bone nodule formation was studied in this in vitro system by direct examination under an inverted phase contrast microscope. The initial event observed was the appearance of cells with phosphatase alkaline activity arranged in several layers and forming a three-dimensional organization around the coral particles. By Day 7, nodule formation began and a refringent material appeared and extended to the background cells during the following days. By Day 15, some coral granules were embedded in a mineralized matrix. Histologic results demonstrated the formation of a mineralized tissue with the appearance of woven bone. This work was supported by the University of Paris VII and the Dental Association of Garancière. This work is a part of Doctorat d'Université that J. M. Sautier will be submitting to the University of Paris VII.     

可诱导结瘤基因nodA的体外转录

报道结瘤基因nodA的体外诱导转录 :当转录体系中有一定量的调控蛋白NodD和植物诱导分子柚皮素 (naringenin )存在时 ,在体外转录的产物中始终存在着与体内转录相一致的特异产物。体外诱导转录表明 ,可诱导结瘤基因nodA在体内转录时 ,可能同样存在NodD蛋白、柚皮素诱导剂分子和nodA启动子的某种方式的直接相互作用     

Heritable variability induced by the in vitro culture and genetic improvement of cultivated plants

Abstract

Genetic variability is found among plants derived from in vitro cultures of somatic cells. A number of different factors, such as the pre-existing genetic variation developed in vivo during tissue differentiation, the variation induced during the in vitro culture and also the selection for specific genotypes during plant regeneration, are considered as possible causes of the phenomenon.

The nature of the genetic changes induced in somaclones (variation in chromosome number, gross and cryptic chromosomal rearrangements, transposition of genetic elements, gene amplification and somatic gene rearrangements) is also discussed.