Exocortis Disease: Evidence for a New Species of “Infectious” Low Molecular Weight RNA in Plants

THE anomalous properties of the infectious agent of exocortis disease in citrus (CEV), supposedly a virus, have been attributed^1,2 to infectious RNA existing as free nucleic acid. Although this model is consistent with the low sedimentation coefficient and the susceptibility to RNAase, the infectious molecule does not cosediment with either single stranded (ss) or double stranded (ds) RNA markers after equilibrium sedimentation in Cs₂SO₄. Resistance of the 8–16S moiety to inactivation by heat or diethylpyrocarbonate further suggests the presence of ds regions₃. A low molecular weight “infectious” RNA species, undetected in healthy tissue, has been implicated as the causal agent of exocortis disease.     

Phylogeny of flowering plants by the chloroplast genome sequences: in search of a “lucky gene”

One of the most complicated remaining problems of molecular-phylogenetic analysis is choosing an appropriate genome region. In an ideal case, such a region should have two specific properties: (i) results of analysis using this region should be similar to the results of multigene analysis using the maximal number of regions; (ii) this region should be arranged compactly and be significantly shorter than the multigene set. The second condition is necessary to facilitate sequencing and extension of taxons under analysis, the number of which is also crucial for molecular phylogenetic analysis. Such regions have been revealed for some groups of animals and have been designated as "lucky genes". We have carried out a computational experiment on analysis of 41 complete chloroplast genomes of flowering plants aimed at searching for a "lucky gene" for reconstruction of their phylogeny. It is shown that the phylogenetic tree inferred from a combination of translated nucleotide sequences of genes encoding subunits of plastid RNA polymerase is closest to the tree constructed using all protein coding sites of the chloroplast genome. The only node for which a contradiction is observed is unstable according to the different type analyses. For all the other genes or their combinations, the coincidence is significantly worse. The RNA polymerase genes are compactly arranged in the genome and are fourfold shorter than the total length of protein coding genes used for phylogenetic analysis. The combination of all necessary features makes this group of genes main candidates for the role of "lucky gene" in studying phylogeny of flowering plants.     

Postactivation Effect in the Deltoid Muscle of Healthy Young Subjects after a Short-Term “Dry” Immersion

Human Physiology - The objective of this study was to characterize, using surface electromyography (EMG), the postactivation effect (PAE) arising in the deltoid muscles of healthy young subjects...     

Light and Electron Microscopic Studies of “Myogranules” in a Child with Hypotonia and Muscle Weakness

Examination by light and electron microscopy of more than 100 muscle biopsies revealed one very unusual case. A 4-year-old boy with non-progressive muscle weakness and hypotonia was found to have small particles, termed “myogranules”, in many muscle fibres from two gastrocnemius biopsies. Paraffin sections and thin sections of plastic-embedded muscle showed that the rod-shaped myogranules measured between 0.1 and 5 microns in length, and were usually orientated in the long axis of the fibre. Normal cross-striations could not be seen in areas occupied by myogranules, although adjacent parts of the same fibre were normal. Electron micrographs showed myofilaments running through the myogranules and a periodicity similar to sections of recrystallized muscle protein paramyosin. It is possible that this child has a disturbance of muscle proteins.     

“Egg transformation” induced by irradiated pollen in Nicotiana: a re-examination

Summary We have attempted to confirm previous reports of egg transformation in seven Nicotiana species, including genetic stocks obtained from the original experimenter. The methods employed in the original experiments were duplicated as closely as possible. In total, 1,622 pollinations were made involving irradiated pollen and mixtures of irradiated and self pollen. Consequently, 995 seedlings from 9,052 seeds were screened for 1,594 potentially detectable transformation events. A very low frequency of unexpected progeny resulted, but these results were not repeatable and appear to have arisen by mechanisms other than transformation. These results are strongly at odds with previous claims, when 50% of offspring were found to be transformed. We conclude that the previous observations of high frequency egg transformation are not reproducible. However, due to the ambiguous nature of the markers employed, it is very difficult to prove that such transformation events do not occur as very rare events.     

Predominance of Single Prophage Carrying a CRISPR/cas System in “Candidatus Liberibacter asiaticus” Strains in Southern China

“Candidatus Liberibacter asiaticus” (CLas) is an uncultureable α-proteobacterium associated with citrus Huanglongbing (HLB, yellow shoot disease), a highly destructive disease affecting citrus production worldwide. HLB was observed in Guangdong Province of China over a hundred years ago and remains endemic there. Little is known about CLas biology due to its uncultureable nature. This study began with the genome sequence analysis of CLas Strain A4 from Guangdong in the prophage region. Within the two currently known prophage types, Type 1 (SC1-like) and Type 2 (SC2-like), A4 genome contained only a Type 2 prophage, CGdP2, namely. An analysis on CLas strains collected in Guangdong showed that Type 2 prophage dominated the bacterial population (82.6%, 71/86). An extended survey covering five provinces in southern China also revealed the predominance of single prophage (Type 1 or Type 2) in the CLas population (90.4%, 169/187). CLas strains with two and no prophage types accounted for 7.2% and 2.8%, respectively. In silico analyses on CGdP2 identified a CRISPR (clustered regularly interspaced short palindromic repeats)/cas (CRISPR-associated protein genes) system, consisting of four 22 bp repeats, three 23 bp spacers and 9 predicted cas. Similar CRISPR/cas systems were detected in all 10 published CLas prophages as well as 13 CLas field strains in southern China. Both Type 1 and Type 2 prophages shared almost identical sequences in spacer 1 and 3 but not spacer 2. Considering that the function of a CRISPR/cas system was to destroy invading DNA, it was hypothesized that a pre-established CLas prophage could use its CRISPR/cas system guided by spacer 1 and/or 3 to defeat the invasion of the other phage/prophage. This hypothesis explained the predominance of single prophage type in the CLas population in southern China. This is the first report of CRISPR/cas system in the “Ca. Liberibacter” genera.     

Premature Mortality from Cardiovascular Disease in the Americas – Will the Goal of a Decline of “25% by 2025” be Met?

Background

Cardiovascular diseases (CVD) are the underlying cause 1.6 million deaths per year in the Americas, accounting for 30% of total mortality and 38% of by non-communicable deaths diseases (NCDs). A 25% reduction in premature mortality due four main NCDs was targeted by the 2011 High-level Meeting of the General Assembly on the Prevention and Control of NCDs. While overall CVD mortality fell in the Americas during the past decade, trends in premature CVD mortality during the same period have not been described, particularly in the countries of Latin America and the Caribbean.

Methods

This is a population-based trend-series study based on a total of 6,133,666 deaths to describe the trends and characteristics of premature mortality due to CVD and to estimates of the average annual percentage of change during the period 2000–2010 in the Americas.

Findings

Premature mortality due to CVD in the Americas fell by 21% in the period 2000–2010 with a -2.5% average annual rate of change in the last 5 year—a statistically significant reduction of mortality—. Mortality from ischemic diseases, declined by 25% - 24% among men and 26% among women. Cerebrovascular diseases declined by 27% -26% among men and 28% among women. Guyana, Trinidad and Tobago, the Dominican Republic, Bahamas, and Brazil had CVD premature mortality rates over 200 per 100,000 population, while the average for the Region was 132.7. US and Canada will meet the 25% reduction target before 2025. Mexico, Costa Rica, Venezuela, Dominican Republic, Panama, Guyana, and El Salvador did not significantly reduce premature mortality among men and Guyana, the Dominican Republic, and Panama did not achieve the required annual reduction in women.

Conclusions

Trends in premature mortality due to CVD observed in last decade in the Americas would indicate that if these trends continue, the Region as a whole and a majority of its countries will be able to reach the goal of a 25% relative reduction in premature mortality even before 2025.     

Regulation of Autophagy by α1-Antitrypsin: “A Foe of a Foe Is a Friend”

Autophagy is involved in both the cell protective and the cell death process but its mechanism is largely unknown. The present work unravels a novel intracellular mechanism by which the serpin α₁-antitrypsin (AAT) acts as a novel negative regulator of autophagic cell death. For the first time, the role of intracellularly synthesized AAT, other than in liver cells, is demonstrated. Autophagic cell death was induced by N-α-tosyl-l-phenylalanine chloromethyl ketone (TPCK) and tamoxifen. By utilizing a fluorescently tagged TPCK analog, AAT was “fished out” (pulled out) as a TPCK intracellular protein target. The interaction was further verified by competition binding experiments. Both inducers caused downregulation of AAT expression associated with activation of trypsin-like proteases. Furthermore, silencing AAT by siRNA induced autophagic cell death. Moreover, AAT administration to cultured cells prevented autophagic cell death. This new mechanism could have implications in the treatment of diseases by the regulation of AAT levels in which autophagy has a detrimental function. Furthermore, the results imply that the high synthesis of endogenous AAT by cancer cells could provide a novel resistance mechanism of cancer against autophagic cell death.     

The Emergence and Evolution of Life in a “Fatty Acid World” Based on Quantum Mechanics

Quantum mechanical based electron correlation interactions among molecules are the source of the weak hydrogen and Van der Waals bonds that are critical to the self-assembly of artificial fatty acid micelles. Life on Earth or elsewhere could have emerged in the form of self-reproducing photoactive fatty acid micelles, which gradually evolved into nucleotide-containing micelles due to the enhanced ability of nucleotide-coupled sensitizer molecules to absorb visible light. Comparison of the calculated absorption spectra of micelles with and without nucleotides confirmed this idea and supports the idea of the emergence and evolution of nucleotides in minimal cells of a so-called Fatty Acid World. Furthermore, the nucleotide-caused wavelength shift and broadening of the absorption pattern potentially gives these molecules an additional valuable role, other than a purely genetic one in the early stages of the development of life. From the information theory point of view, the nucleotide sequences in such micelles carry positional information providing better electron transport along the nucleotide-sensitizer chain and, in addition, providing complimentary copies of that information for the next generation. Nucleotide sequences, which in the first period of evolution of fatty acid molecules were useful just for better absorbance of the light in the longer wavelength region, later in the PNA or RNA World, took on the role of genetic information storage.     

Neuronal Expression of Splice Variants of “Glial” Glutamate Transporters in Brains Afflicted by Alzheimer’s Disease: Unmasking an Intrinsic Neuronal Property

Anomalies in glutamate homeostasis may contribute to the pathological processes involved in Alzheimer’s disease (AD). Glutamate released from neurons or glial cells is normally rapidly cleared by glutamate transporters, most of which are expressed at the protein level by glial cells. However, in some patho-physiological situations, expression of glutamate transporters that are normally considered to be glial types, appears to be evoked in populations of distressed neurons. This study analysed the expression of exon-skipping forms of the three predominant excitatory amino acid (glutamate) transporters (EAATs1-3) in brains afflicted with AD. We demonstrate by immunocytochemistry in temporal cortex, the expression of these proteins particularly in limited subsets of neurons, some of which appeared to be dys-morphic. Whilst the neuronal expression of the “glial” glutamate transporters EAAT1 and EAAT2 is frequently considered to represent the abnormal and ectopic expression of such transporters, we suggest this may be a misinterpretation, since neurons such as cortical pyramidal cells normally express abundant mRNA for these EAATs (but little if any EAAT protein expression). We hypothesize instead that distressed neurons in the AD brain can turn on the translation of pre-existent mRNA pools, or suppress the degradation of alternately spliced glutamate transporter protein, leading to the “unmasking” of, rather than evoked expression of “glial” glutamate transporters in stressed neurons. Special issue article in honor of Dr. Graham Johnston.     

Dispersal of non-myrmecochorous plants by a “keystone disperser” ant in a Mediterranean habitat reveals asymmetric interdependence

In contrast to other plant–animal mutualisms, seed dispersal interactions, and particularly seed dispersal by ants, are generally considered asymmetric, non-specialized relationships in which dispersers depend less on plants than vice versa. Although myrmecochory is well understood in many terrestrial ecosystems, dispersal of non-elaiosome-bearing seeds by ants has barely been studied outside the Neotropics. Aphaenogaster senilis, a common ant in Southern Spain, collects a great variety of non-myrmecochorous diaspores along with insect prey. At our study site, fleshy fruits of Arum italicum, Phillyrea angustifolia and Pistacia lentiscus represent up to one-fourth of the items collected by A. senilis from June to November. However, they are mostly ignored by other ants. In the laboratory, the addition of A. italicum fruits to A. senilis insect-based diet increased male production and both worker and queen pupae size. Seeds were transported up to 8 m away from the mother plant and deposited in a favorable habitat allowing a relatively high proportion of germination. Given important differences in seed production between species, our data suggest that A. senilis removes virtually all seeds of A. italicum, but a negligible fraction of P. lentiscus seeds. We conclude that in contrast to the common view, dispersal of non-myrmecochorous Mediterranean plants by ants might be an important phenomenon. Keystone disperser ants like A. senilis probably obtain an important fitness advantage from non-myrmecochorous diaspore collection. However, plant benefit may vary greatly according to the amount of seeds per individual plant and the existence of alternative dispersal agents.     

A Review of “Maritime Power and the Law of the Sea: Expeditionary Operations in World Politics”

Abstract

The purpose of the London Dumping Convention (LDC) is the protection of the marine environment including its seabed and subsoil. The preamble, articles, and annexes of the LDC make clear that the Convention must be interpreted in a manner which ensures that this responsibility is met. As a partial response to that mandate, the LDC prohibits the dumping at sea of certain wastes, including high‐level radioactive wastes. Disposal of high‐level wastes in seabed sediments is the subject of ongoing technical, environmental, and engineering feasibility studies by several countries. In the LDC's definition of dumping, the phrase “disposal at sea”; could be interpreted narrowly to mean the final resting place of wastes—with seabed disposal excluded from coverage because those wastes are not in direct contact with “marine waters.”; Given the LDC's object and purpose, though, the only harmonious and reasonable interpretation is that which defines “disposal at sea”; to mean the place where the dumping activities occur. Other international agreements also support this object and purpose‐based interpretation which concludes that seabed disposal is covered and prohibited. In addition, this approach is preferred because it contributes to the continued effectiveness of the LDC.     

An Exploration of Computer Game-Based Instruction in the “World History” Class in Secondary Education: A Comparative Study in China

So far, many studies on educational games have been carried out in America and Europe. Very few related empirical studies, however, have been conducted in China. This study, combining both quantitative with qualitative research methods, possibly compensated for this regret. The study compared data collected from two randomly selected classes (out of 13 classes) under computer game-based instruction (CGBI) and non-computer game-based instruction (NCGBI), respectively, in a senior high school located in Nanjing, Capital of Jiangsu Province, in China. The participants were 103 students, composed of 52 boys and 51 girls (aged 17-18 years old). The following conclusion was reached: (1) participants under CGBI obtained significantly greater learning achievement than those under NCGBI; (2) participants were significantly more motivated by CGBI compared with NCGBI; (3) there were no significant differences in learning achievement between boys and girls; although (4) boys were significantly more motivated by CGBI than girls. Both disadvantages and advantages were discussed, together with directions for future research.     

Growth of a photosynthetic bacterium anaerobically in darkness,supported by “oxidant-dependent” sugar fermentation

Rhodopseudomonas capsulata can obtain energy for growth from light (anaerobically) and can also grow heterotrophically in darkness using alternative energy conversion modes, namely, aerobic respiration or an unusual type of anaerobic catabolism of sugars. Dark anaerobic growth with fructose as sole carbon and energy source is dependent on the presence of an accessory oxidant such as trimethylamine-N-oxide, is accompanied by production of lactate and other classical fermentation products, and yields cells with a high content of photosynthetic pigments and polyhydroxybutyrate.Abbreviation Used TMAO trimethylamine-N-oxide - DMSO dimethyl sulfoxide - EMP Embden-Meyerhof-Parnas - ED Entner-Doudoroff - PHB polyhydroxybutyrate