Distinctive patterns of translational reinitiation in the lac repressor mRNA: bridging of long distances by out-of-frame translation and RNA secondary structure, effects of primary sequence.

In the early region of the Escherichia coli lac repressor mRNA, translational reinitiation events triggered by nonsense codons occur over long distances and in a distinctive pattern not explained by simple use of the next available initiator triplet. Defined fusions of the restart sites to the lacZ coding region have been used to explore the basis for these reinitiation patterns and to ask whether the sites can function in independent initiation at the 5' end of an mRNA. The results obtained confirm earlier indications that the restart sites may have little or no inherent capacity for binding free 30S ribosomes. The data also add to growing evidence that primary sequence elements are important determinants of reinitiation efficiency. On the basis of the reinitiation activities for nonsense sites throughout the early region of the mRNA, we suggest that out-of-frame restarts and RNA secondary structure bridge long distances between the point of termination and downstream restart codons. Such bridging mechanisms could serve more generally as a means of propagating translational activity across long polycistronic mRNAs.     

Localization of calcium in skeletal and cardiac muscle

Summary The requirement of calcium (Ca²⁺) in the excitation-contraction coupling of both skeletal and cardiac muscle is well established. However, the exact location of the intracellular storage sites of Ca²⁺ is not firmly established. We report here on the ultrastructural distribution of Ca²⁺ in white and red skeletal muscle and in cardiac muscle of the rat using combined phosphate-pyroantimonate (PPA) and oxalate-pyroantimonate (OPA) procedures. The methods are based on (a) stabilization and/or trapping of Ca²⁺ during the primary fixation step in glutaraldehyde by potassium phosphate or oxalate; (b) subsequent wash-out of all non-trapped cations such as Na⁺ and Mg²⁺ in potassium phosphate or oxalate; (c) conversion of the complexed or trapped Ca²⁺ into an electron-dense calcium pyroantimonate salt in 100 m-thick tissue sections; and (d) wash-out of the excess potassium pyroantimonate at alkaline pH.With the OPA procedure, mitochondria of all muscle types showed little precipitate. The junctional sarcoplasmic reticulum was stongly reactive in relaxed white skeletal muscle, negative in contracted white fibres and negative in red skeletal and cardiac muscle, independent of the state of relaxation-contraction. Other organelles were essentially free of deposits.With the PPA method, the precipitate was almost exclusively confined to the sarcolemma and its T-tubular invaginations in cardiac and slow skeletal muscle, and was absent in fast skeletal muscle. Apart from occasional deposits in mitochondria, all other organelles were free of precipitate. The sarcolemma-associated deposits were clearly confined to the inner leaflet of the lipid bilayer. The amount of precipitate varied within the contraction cycle, relaxed cells possessing the highest density.Exposure of the tissue to La³⁺ resulted in the complete absence of sarcolemma-bound precipitate suggesting that the Ca²⁺ is exchangeable. Furthermore, these cytological data suggest a basic difference in Ca²⁺ storage between white skeletal muscle on the one hand, and red skeletal and cardiac muscle on the other.     

Localization of GABA-like immunoreactivity in the ectostriatum of domestic chicks: GABA immunocytochemistry combined with Golgi impregnation

Brain Cell Biology - The distribution of GABA-like immunoreactivity (GABA-LI) in the ectostriatal core (Ec) of domestic chicks (one to two days old) was investigated using (1) preembedding GABA...     

Translation of phage f1 gene VII occurs from an inherently defective initiation site made functional by coupling

Expression of the filamentous phage f1 gene VII is shown to be translationally coupled to that of the upstream gene V. Fusions of the gene VII initiation site to the lacZ coding region were used to determine that initiation at the VII site is completely dependent on the process of translation having proceeded up to a stop codon immediately upstream from the VII site. Coupled expression from the VII site was found to be inefficient, proportional to the level of upstream translation, and very sensitive to the distance from the functional upstream stop codon. Independent expression from the VII site was not observed, even in a deletion series designed to remove potentially masking RNA structure. On the basis of the VII site's dissimilarity to ribosome binding site sequences and its properties overall, we suggest that it inherently lacks the features required for independent recognition by ribosomes, and acquires the ability to initiate synthesis of gene VII protein by virtue of the coupling process.     

Mapping of two new markers within the smallest interval harboring the spinal muscular atrophy locus by family and radiation hybrid analysis

The locus responsible for the childhood-onset proximal spinal muscular atrophies (SMA) has recently been mapped to an area of 2–3 Mb in the region q12–13.3 of chromosome 5. We have used a series of radiation hybrids (RHs) containing distinct parts of the SMA region as defined by reference markers. A cosmid library was constructed from one RH. Thirteen clones were isolated and five of these were mapped within the SMA region. Both RH mapping and fluorescence in situ hybridization analysis showed that two clones map in the region between loci D5S125 and D5S351. One of the cosmids contains expressed sequences. Polymorphic dinucleotide repeats were identified in both clones and used for segregation analysis of key recombinant SMA families. One recombination between the SMA locus and the new marker 9Ic (D5S685) indicates that 9Ic is probably the closest distal marker. The absence of recombination between the SMA locus and marker Fc (D5S684) suggests that Fc is located close to the disease gene. These new loci should refine linkage analysis in SMA family studies and may facilitate the isolation of the disease gene.     

Tropical rain forest types and soil factors in a watershed area in Guyana

Abstract. An inventory was made of the vegetation and soils of a watershed area of 480 ha in the tropical rain forest of Guyana. 252 plots of 0.05 ha were sampled. In total 111 tree species > 20 cm were recorded. A TWINSPAN analysis resulted in seven groupings. Correspondence Analysis revealed that the major environmental differentiation underlying the floristic variation is according to soil type. Finally, five main forest communities were described. Most sites in the watershed area are characterized by the dominance of one or a few species. Species distribution patterns are strongly determined by soil type and drainage class.     

The impact of acidification on diatoms and chemistry of Dutch moorland pools

Summary Moorland pools are shallow oligotrophic soft water lakes on poorly buffered sandy soils. Diatom assemblages of samples from 16 pools taken in 1920 and 1978 were compared by analysis of pH-spectra, diversity, dissimilarity and multivariate statistical techniques.The pH-spectra of pools in the southern (S) and central (C) part of the country indicate a fall in pH from 4.5–6.0 in the old samples to 3.7–4.6 in the recent ones. The pH-spectra of the northern pools (N) do not indicate a significant shift from the original pH (ca 4.5).The number of species in the count and the diversity (indices of Simpson and Shannon) decreased significantly in S+C, and that goes also for the dissimilarity index of Dyer. No changes were found in N.The first component (PC 1) of the principal component analysis explains 61% of total variance. PC 1 is correlated with log [SO₄] (r=0.83, p<0.001) and even better (r=0.95, p<0.001) with the relative sulphate concentration,i.e. the ratio of sulphate to all major anions (sulphate, chloride, bicarbonate). All old samples have low scores on PC 1, recent samples have low scores on the second (PC 2) and third (PC 3) principal component. Old samples have high scores on PC 2 and PC 3, explaining 9 and 6% of total variance, respectively.The orginal variation, caused by regional factors, is replaced by a SO₄ ^2– controlled variation. PC 1 is nearly completely determined by the relative abundance ofEunotia exigua. This species, which is known to be very resistant to pollution by sulphur, aluminium and heavy metals, increased largely from 1920 to 1978.In spite of the rather homogeneous distribution of wet sulphate deposition in the Netherlands, substantial differences in SO₄ ^2– content in the pools are observed, being lowest in N (0.13–0.48 meq.l^–1) and highest in S+C (0.38–1.65 meq.l^–1). Sulphate is positively correlated with calcium, aluminum and magnesium but negatively with factors that characterize humic acid waters (e.g. permanganate-consumption, iron and the ratio of univalent to divalent cations). Sulphate concentration depends on the intensity of sulphate reduction, accumulation by dry deposition in surrounding forests of Scots pine, drought and atmospheric deposition.     

Sensitivity to weighting in life cycle impact assessment (LCIA)

The International Journal of Life Cycle Assessment - Weighting in life cycle assessment (LCA) incorporates stakeholder preferences in the decision-making process of comparative LCAs. Research...     

Synthesis and Biological Activity of Some 2-Aminopurine Carbonucleosides

Abstract

A series of new o ne t wo subtituted c arbonucleoside analogues (OTC) of purine was synthesized and evaluated against cytomegalovims and varicella-zoster virus in human embryonic lung (HEL) cells.     

INFLUENCE OF FRESHWATER FLOW REGIME ON FISH ASSEMBLAGES IN THE GREAT FISH RIVER AND ESTUARY

Summary

Two and a half years of data were collected from the lower Great Fish River, head region and estuary to determine the fish species composition within these areas. Gilchristella aestuaria, Liza dumerilii, Rhabdosargus holubi and Pomadasys commersonnii were the four most abundant species captured, with riverine flow rate having an important effect on both species composition and numbers of fishes in the different regions. Most marine species displayed a strong inverse relationship between catch per unit effort and elevated freshwater inputs. Euryhaline marine species dominated the catches at all sampling sites during low flows but were less common during high flow periods when catadromous species were most abundant. Based on the available evidence it is suggested that for most marine species in the river this decline in abundance is related to low conductivity levels following floods rather than avoidance of elevated flows. The impact of elevated suspensoid concentrations and lowered dissolved oxygen concentrations on freshwater and estuarine fish populations during major river flooding is also discussed.