Isolation of intact polysomes from mechanically dehulled developing grain

A rapid method for obtaining large quantities of developing groats suitable for the isolation of highly intact polysomes has been developed. Developing spikelets were harvested directly from oat panicles into liquid nitrogen and then quickly passed through a dehuller. Chaff was removed by air aspiration and the resultant groats were collected directly back into liquid nitrogen. Approximately 250 g of groats could be isolated each man-hour by the above method. In comparison, only 10 g of endosperm could be collected by squeezing it out of spikelets using an endosperm mangle. Membrane-bound polysomes extracted from the immature groats were compared to those extracted from endosperm. The largest polysomes discernable as unique peaks on sucrose gradients were ten-mers and nine-mers for groats and endosperm, respectively. Polysomes isolated from both starting materials stimulated similar incorporations of [³⁵S]methionine into trichloroacetic acid-insoluble products during in vitro translations in wheat germ extract. Both polysome preparations directed the synthesis of similar high-molecular-weight proteins. Based on these criteria, polysomes from both preparations were found to be of similar intactness, although the groat starting material was much more readily obtained. The polysome classes having the maximum absorbance peak for endosperm and groat polysomes were six-mers and eight-mers, respectively.     

PHOTOSYNTHETIC QUANTUM EFFICIENCIES OF PHYTOPLANKTON FROM PERENNIALLY ICE COVERED ANTARCTIC LAKES1

A recently introduced approach far estimating the photosynthetic quantum efficiency (φ) of a freshwater or marine phytoplankton community has been applied for the first time to high latitude polar ecosystems, namely four lakes of southern Victoria Land, Antarctica. Values for φ at various depths ranged from 0.0022–0.1560 when calculated using a recommended mean extinction coefficient for phytoplankton (i.e. k?_c= 0.016). By contrast, φ ranged from 0.0037–0.0760 when calculated using an empirically estimated value for k?_c of 0.0328. If the recommended k?_c= 0.016 more closely approaches an accurate estimate, then the φ valves indicate that the phytoplankton convert light to organic carbon more efficiently than elsewhere. However, if the empirically derived k?_c= 0.0328 more closely approaches an accurate estimate, then the φ values indicate the phytoplankton trap light more efficiently than elsewhere. Although we have not resolved whether light conversion (φ) or light trapping are more efficient, the results show that the phytoplankton of these Antarctic lakes are well adapted to performing photosynthesis under extremely low light conditions.     

A simple sensitive radioreceptor assay for calcium antagonist drugs

A radioreceptor assay for calcium channel antagonist drugs described here is based on the ability of these drugs to affect ³H-nitrendipine binding to calcium channels. All the known calcium channel antagonists may be assayed in this manner. The assay can detect 10–100 nM (4 – 40 ng/ml) nimodipine, 10–100 nM (3.5 – 35 ng/ml) nifedipine, 3–30 μM (1.2 – 12 μm/ml) prenylamine, 0.1 – 1.0 μM (49 – 490 ng/ml) verapamil and 3–30 μM (1.2 – 12 μg/ml) diltiazem. These values cover the range of concentrations of calcium channel antagonists that are clinically important. As the radioreceptor assay detects active metabolites as well as the parent drugs, it should prove a useful adjunct in cardiovascular therapy. The method is more reproducible, simpler and less expensive than other methods such as high pressure liquid chromatography.     

Haemagglutinin activity in the haemolymph of individual Acrididae (grasshopper) specimens

Twenty-four individual grasshopper specimens representing four Melanoplus spp. contained similar broad-spectrum haemolymphatic haemagglutinin. The agglutinin activity showed highest titre toward human ABO and rabbit cells among nine types of erythrocytes tested. Titre values differed between individual insects but agglutination specificity toward different erythrocytes was similar. Agglutination of type-O red cells by individual grasshopper haemolymph was inhibited by 34 of 41 tested carbohydrates, carbohydrate derivatives, alcohols and chelating agents. Individual insects showed similar patterns of haemagglutination inhibition. Non-inhibitory compounds were mannose and mannose derivatives (excepting N-acetylneuraminate), several glucose derivatives, amino sugars and ethanol. The observations indicated that haemolymph from an individual grasshopper contained complex heteroagglutinin activity similar to that found in haemolymph pooled from several insects. Determination of minimal effective inhibitor concentrations confirmed the presence of heteroagglutinin activity primarily directed toward galactose and glucose and related α-linked glycosidic derivatives.     

Proliferation of cells undergoing chondrogenesis in vitro

Abstract. Continuous exposure of chicken embryo limb bud mesenchyme cells undergoing chondrogenesis in vitro to [³H] thymidine ([³H]TdR) revealed that more than 90% of the cells synthesized DNA at least once during 120 h of culture. When cells were exposed to [³H]TdR for 24 h beginning at various times throughout the culture period, the percentage of cells which incorporated [³H]TdR during each period was approximately 92%. However, when the period for incorporation of radioisotope was limited to two hours, the number of cells which incorporated [³H]TdR was found to decline during chondrogenesis in vitro. This decline was coincident with the appearance of extracellular matrix material and occurred in those cells which had, and had not, expressed the cartilage phenotype.
We conclude from these studies that (1) practically all of the cells continue to proliferate while chondrogenesis is occurring in vitro, (2) there is an increase in the length of the cell cycle during chondrogenesis in vitro, and (3) withdrawal from the cell cycle is not required for differentiation of mesenchyme into cartilage.     

DNA repair kinetics in irradiated undifferentiated and terminally differentiated cells

The brains of male Fisher 344 rats bearing 80-150 mg intracerebral 9L/Ro tumors were irradiated with doses of 1,250-5,000 rads of x- or gamma-rays. At various times after irradiation, the cerebellum and tumor were excised, dissociated into single cells and the DNA from these cells sedimented through alkaline sucrose gradients in zonal rotors with slow gradient reorienting capability. Quantitation of the DNA repair kinetics demonstrated that the process in both tumor cells and neurons has a fast and slow phase. Although all other alternatives cannot be completely eliminated, we suggest that these two phases are most reasonably interpreted as representing repair of lesions in very accessible and less accessible regions of the genome rather than 1) repair of different types of lesions such as single- or double-strand breaks or 2) removal of immediate breaks and breaks induced during excision repair of latent base damage. The slow repair phase is saturable, but not inducible in both tumor cells and neurons. The data suggest that tumor cells restore their chromosomal DNA structure to the unirradiated state faster than neurons because 1) they contain more of the repair system per unit of DNA and 2) a larger proportion of their genetic material is comprised of very accessible regions. The data also suggest that the entire tumor cell genome may be accessible to the repair enzyme(s), while it is possible that a portion of the neuronal genome may be completely inaccessible.     

Unprocessed Bran in Treatment of Diverticular Disease of the Colon

Seventy patients with diverticular disease of the colon were treated with a high-residue, low-sugar diet including unprocessed bran. Follow-up for an average of 22 months showed marked relief of symptoms in 62 patients. Bowel habit was restored towards normal and abdominal discomfort relieved. Only seven patients were unable to give up the use of laxatives. Eight patients did not tolerate the bran diet, and one of these needed surgical treatment. None of the 62 patients who took the diet needed surgery.     

Transformation of Murine Cells by Two “Slow Viruses,” Visna Virus and Progressive Pneumonia Virus

Visna and progressive pneumonia virus (PPV), two antigenically related, non-oncogenic "slow viruses" which have ribonucleic acid (RNA)-dependent deoxyribonucleic acid (DNA) polymerase activity, were examined for their ability to transform cells. Murine cells which had been exposed to either visna or PPV developed foci of altered, spindle-shaped cells 3 to 4 weeks after infection. Visna and PPV transformed lines were established from these cultures. There was no evidence that other oncogenic DNA or RNA viruses were involved in the observed transformation. Visna or PPV could be "rescued" from all transformed lines by co-cultivation with normal sheep testis cells. "Rescued" virus was identified as visna or PPV, and they retained the capacity to transform mouse cells. These experiments may have important implications in the understanding of both viral carcinogenesis and "slow" viral infections.     

CYTOKINESIS AND PLASMODESMATA IN ULOTHRIX1

Cytokinesis essentially similar to that of vascular plants occurs in Ulothrix, an unbranched filamentous green alga. Plasmodesmata, similar to those of vascular plants, but different from those of many other algae, are also present. Cell plate formation and plasmodesmata also occur in Stigeoclonium, a branched green alga.