Length heterogeneity of the large spacer of Vicia faba rDNA is due to the differing number of a 325 bp repetitive sequence elements

Summary The DNA fragments including the whole large spacer region of Vicia faba rDNA were cloned in plasmid pBR325. Sixteen clones were classed into five groups which differed from each other in the lengths of the rDNA inserts. Physical maps of these length variants cloned were constructed using EcoRI, SalI, HpaI, MluI and AccI and evidence was obtained that the length heterogeneity was due mainly to the differing number of 325 base pairs (bp) subrepeating elements in the large spacer. Sequence analysis of this subrepeating element revealed that it consisted of a duplet of an approximately 155 bp sequence and a 14 bp unrelated sequence. This structure of the repetitive element is novel.     

Correlations in one-dimensional fully developed chaos

Correlations in the baker map and the tent map as examples of one-dimensional, fully developed chaos are considered. It is shown, utilizing symbolic dynamical systems derived from these maps, that the vanishing second-order correlation function is not sufficient to guarantee uncorrelatedness. Importance of the higher-order, especially third-order, correlation functions is emphasized for chaotic systems. In search of the quantities that grasp correlational behaviors as a whole in chaotic systems, it is proposed to use the fixed-separation correlation integral, which is a modified quantity of the usual correlation integral devised to calculate the fractal dimension of strange attractors, for these maps. It is shown that the new quantity contains all the even-number orders of autocorrelation function that are commonly considered.     

Structural changes of alpha2- and ovomacroglobulins

The plasma α₂-macroglobulin and its egg white homologue ovomacroglobulin were purified from several different species and their structure before and after the reaction with proteinases studied by electron microscopy. The negatively stained specimens showed either a ringlike structure or a flowerlike one before the reaction with proteinses, but their structures changed into open rectangular ones after the reaction. The translational frictional ratio f/f ₀ of human α2-macroglobulin and crocodilian ovomacroglobulin given in the literature is between 1.5 and 1.6 before and after the reaction with proteinases. The value reflects asymmetry due not to a high axial ratio, but rather to an openness of the structure resulting in a partially free draining character of the molecules. The computational method developed by Bloomfield and his co-workers based on the formalism of Kirkwood is used to calculate the frictional ratio of several models constructed from small spheres. The overall shape of the models is derived from electron micrographs. Although the degree of hydration is an unknown parameter in the calculation, reasonable agreement is obtained between the experimental values of f/f ₀ and the calculated ones. Combination of electron microscopic and hydrodynamic methods would be fruitful in the structural study of giant proteins such as α₂-macroglobulin.     

Cytoplasmic effects on the tissue culture response of wheat (Triticum aestivum) callus

Summary Calli were initiated from immature embryos of eight lines of hexaploid wheat (Triticum aestivum L. em. Thell) with different cytoplasms, the euplasmic nuclear donor Chinese Spring and seven alloplasmic lines derived from wild relative species of the genera Triticum and Aegilops. The calli were found to differ in their initial growth rates, their sensitivity to 2,4-D and their ability to organise shoot primordia, demonstrating that the cytoplasm can significantly affect the behaviour of tissues in culture. The potential for improving the responses of tissues in culture by cytoplasmic changes is noted.     

Responses of aerial ventilation to hypoxia and hypercapnia inChanna argus,an air-breathing fish

Summary The snake-head fish (Channa argus) is an obligate air-breather inhabiting fresh waters in the temperate zone of East Asia.Ventilation of the air-breathing organ and aerial gas exchange were measured in 1 to 2 kg specimens at 15 and 25°C. Additionally, the ventilatory responses to hypoxia and hypercapnia were studied. Aerial ventilation increased from 1.1 to 2.9 mlbtps·kg^–1·min^–1 when temperature rose from 15 to 25°C. Concomitantly, O₂-uptake through airbreathing increased from 0.1 mlstpd·kg^–1·min^–1 (15°C) to 0.28 mlstpd·kg^–1·min^–1 (25°C), whereas aerial gas exchange was less important for CO₂-climination as evident from low gas exchange ratios (0.16 at 15°C, 0.29 at 25°C).Ventilation increases only slightly in response to inspiration of hypercapnic gas mixtures or to hypoxic conditions in water. By contrast, inspiration of hypoxic gas mixtures caused marked increases of ventilation in particular at the higher temperature.Aerial ventilation inChanna is low compared to values for ectothermic pulmonary breathers. However, its ventilatory responses to hypoxia strikingly resemble those of reptiles: The most marked ventilatory response to hypoxia occurs at the higher temperature where the demands for O₂ are greatest.     

Hormonal regulation of berberine production in cell suspension cultures of Thalictrum minus

Effects of auxin and cytokinin on cell growth and alkaloid production in cell suspension cultures of Thalictrum minus were examined in an attempt to increase the productivity of a medicinal compound, berberine. In Linsmaier and Skoog medium containing auxin such as 2,4-D (1 M), the cultured cells grew rapidly, producing little berberine. On the other hand, the berberine-producing activity was remarkably enhanced by simultaneous administration of auxin and cytokinin, although cell growth was inferior. In particular, for the combination of NAA (60 M) and 6-benzylaminopurine (10 M), the yield of berberine was as high as 20 mg/30 ml medium after 2 weeks of culture. Furthermore, most of the berberine produced by the cells was released into the liquid medium, in which an excess of berberine crystallized. The results of the present experiments are suggestive of an advantage in adopting a two-stage culture method for the production of berberine in fermentor systems.Abbreviations LS Linsmaier and Skoog - 2,4-D 2,4-dichlorophenoxyacetic acid - NAA 1-naphthaleneacetic acid - IAA indole-3-acetic acid - IBA indole-3-butyric acid - BAP 6-benzylaminopurine     

Enzymatic difference between laticifers and cultured cells of papaya

Cell suspension cultures of Carica papaya were found to produce a papain-like enzyme showing an amidase activity similar to papain. Experiments suggested that the enzyme was an SH-enzyme, but an immunological test indicated the absence of papain in cultured cells. The isoelectric point (4.3) of the enzyme of cultured cells was the same as that of the leaf extract, but was different from that of papain or other amidases in the latex of the fruit.     

Heterogeneity of keratin distribution in the oral mucosa and skin of mammals as determined using monoclonal antibodies

Summary The immunohistochemical localization of keratins in the oral epithelia of several mammals was investigated using the monoclonal antibodies to keratins, PKK1 (41–56 kilodaltons) and KL1 (55–57 kilodaltons). The staining patterns obtained in different locations of the oral mucosa and of the skin epidermis were compared. In the papillae on the dorsal surface of the tongue, some areas exhibited marked PKK1 staining, while other area were PKK1 negative. In general, rodent oral epithelia were negative for PKK1 in the basal layer, while comparatively strong PKK1 staining was observed in cells of the upper spinous layer. In the epidermis, positive PKK1 reactions were confined to the basal layer, while KL1 staining was occasionally seen in the basal layer of oral epithelia. In cats, dogs, and monkeys, different PKK1 and KL1 binding patterns were observed in oral epithelia. Also, the distribution in oral epithelia differed from that seen in the epidermis of these animals. In the epidermis, the distribution of PKK1 and KL1 was regular, with PKK1 usually being confined to the basal layer, while KL1 binding was found in the spinous and granular cell layers, and was dependent on the degree of keratinization. In the animals studies, keratin expression as detected by PKK1 and KL1-was different in the skin epidermis and oral epithelia, and the localization of these keratins differed in the various types of oral mucosa.     

Expression of adenovirus type 12 early region 1 in KB cells transformed by recombinants containing the gene.

The adenovirus type 12 (Ad12) early region 1 (E1) gene was introduced into KB cells by using a dominant selection vector, pSV2-gpt, and over 80 Gpt+ KB cell clones were established. Three types of recombinant DNAs (gAE1A, gARC, and gABA) were constructed. They contained the AccI-H, EcoRI-C, and BamHI-A fragments, respectively, of Ad12 DNA in pSV2-gpt. Five of 50 (10%) gABA-transformed cell clones, 12 of 18 (67%) gAE1A-transformed cell clones, and 10 of 18 (56%) gARC-transformed cell clones complemented the growth of Ad5 dl312 (deletion in E1A) and were designated as Gpt+ Ad+ cell clones. In these cell clones at their early passages, recombinant genome sequences were detected in cellular DNA and were expressed. T antigen g (the E1A gene product) was detected by immunofluorescence. The Gpt+ Ad+ cell clones supported the growth of Ad5 deletion mutants in parallel with the expression of Ad12 E1A or E1A plus E1B genes. After infection of Gpt+ Ad+ cell clones with Ad5 dl312, the early genes of dl312 were efficiently transcribed, indicating the expression of the pre-early function of the Ad12 E1A gene. Two clones each from gAE1A-,gARC-, and gABA-transformed cells were subcultured for a long period to determine the stability of the transfecting DNAs. Subculture in a nonselective medium resulted in cells which lost the transfecting DNAs. Subculture in a selective medium resulted in the selection of cells which maintained the gpt gene expression but lost the Ad12 gene expression. These results indicate that the transfecting DNA is present in an unstable state in KB cells.     

Effects of Some Growth Regulators and Benzoic Acid Derivatives on Flower Initiation and Root Elongation of Pharbitis nil, Strain Kidachi

Seedlings of Pharbitis nil, strain Kidachi, were grown undercontinuous light at 20°C in vessels containing 5,000-mlnutrient solution, 24 plants per vessel. NAA (0.005–0.5µM), GA₃ (0.1–0.5 µM), kinetin (0.5–5µM), benzyladenine (0.05–5 µM) or abscisicacid (4 µM) added to the nutrient solution induced long-dayflowering, and the flowering was always accompanied by suppressionof root elongation. 3,4-Dichlorobenzoic acid (0.05–10µM) and some other benzoic acid derivatives which arehighly effective for the induction of flowering in Lemna paucicostataalso showed similar effects. Neither NAA, kinetin nor 3,4-dichlorobenzoicacid applied via the apical part of the hypocotyl could causeflowering or suppression of root elongation. Thus, the flower-inducingeffect of the above substances was presumed to be secondaryto the suppression of root elongation. Ethrel (1–50 µM)added to the nutrient solution suppressed root elongation, butdid not induce flowering probably because it has flower-inhibitingactivity. ¹ This paper is dedicated to the memory of Dr. Joji Ashida,the first president of the Japanese Society of Plant Physiologists. (Received December 15, 1982; Accepted February 25, 1983)