Current status of Paragonimus and paragonimiasis in Ecuador

A review of national and international publications on paragonimiasis in Ecuador,epidemiological records from the Ministry of Public Health and unpublished researchdata was conducted to summarise the current status of the parasite/disease. Thepurpose of the review is to educate physicians, policy-makers and health providers onthe status of the disease and to stimulate scientific investigators to conductfurther research. Paragonimiasis was first diagnosed in Ecuador 94 years ago and itis endemic to both tropical and subtropical regions in 19 of 24 provinces in thePacific Coast and Amazon regions. Paragonimus mexicanus is the onlyknown species in the country, with the mollusc Aroapyrgus colombiensisand the crabs Moreirocarcinus emarginatus,Hypolobocera chilensis and Hypolobocera aequatorialisbeing the primary and secondary intermediate hosts, respectively. Recentstudies found P. mexicanus metacercariae in Trichodactylusfaxoni crabs of the northern Amazon. Chronic pulmonary paragonimiasis iscommonly misdiagnosed and treated as tuberculosis and although studies havedemonstrated the efficacy of praziquantel and triclabendazole for the treatment ofhuman infections, neither drug is available in Ecuador. Official data recorded from1978-2007 indicate an annual incidence of 85.5 cases throughout the 19 provinces,with an estimated 17.2% of the population at risk of infection. There are no currentdata on the incidence/prevalence of infection, nor is there a national controlprogramme.     

Interaction of Ditylenchus dipsaci and Meloidogyne hapla on Resistant and Susceptible Plant Species

Numbers ofDitylenchus dipsaci or Meloidogyne hapla invading Ranger alfalfa, Tender crop bean, Stone Improved tomato, AH-14 sugarbeet, Yellow sweet clover, and Wasatch wheat from single inoculations were not significantly different from numbers by invasion of combined inoculations. D. dipsaci was recovered only from shoot and M. hapla only from root tissue. Combined inoculations did not affect reproduction of either D. dipsaci or M. hapla. D. dipsaci suppressed shoot growth of all species at 15-30 C, and M. hapla suppressed shoot growth of tomato, sugarbeet, and sweet clover at 20, 25, and 30 C. There was a positive correlation (P < 0.05) between shoot and root growth suppression by D. dipsaci on all cultivars except wheat at 20 C and tomato at 30 C. M. hapla suppressed (P < 0.05) root growth of sugarbeet at 20-50 C and wheat at 30 C. Growth suppression was synergistic in combined inoculations of sweet clover shoot growth at 15 C and root growth at 20-30 C, wheat root growth at 15 and 20 C, and tomato root growth at 15-30 C (P < 0.05) D. dipsaci invasions caused mortality of alfalfa and sweet clover at 15-30 C and sugarbeet at 20-30 C. Mortality rates of alfalfa and sweet clover increased synergistically (P < 0.05) from combined inoculations.     

Host Status of Seven Weed Species and Their Effects on Ditylenchus destructor Infestation of Peanut

The host suitability to Ditylenchus destructor of seven common weed species in peanut (Arachis hypogaea) fields in South Africa was determined. Based on the number of nematodes per root unit, white goosefoot (Chenopodium album), feathertop chloris (Chloris virgata), purple nutsedge (Cyperus rotundus), jimson weed (Datura stramonium), goose grass (Eleusine indica), khaki weed (Tagetes minuta), and cocklebur (Xanthium strumarium) were poor hosts. Ditylenchus destructor survived on all weed species; population densities increased in peanut hulls and caused severe damage to seeds of peanut grown after weeds. Roots of purple nutsedge left in the soil suppressed populations of D. destructor and root and pod development in peanut grown after the weed. However, nematode populations in peanut hulls and seeds were not suppressed. Some weed species, especially purple nutsedge which is common in peanut fields, can be used to indicate the presence of D. destructor in the absence of peanut.     

The melectine bee genera Brachymelecta and Sinomelecta (Hymenoptera,Apidae)

The enigmatic, cleptoparasitic bee genera Brachymelecta Linsley and Sinomelecta Baker (Apinae: Melectini) are redescribed, each represented by a single species which has not been reencountered since capture of the type series ca. 1878 and 1900, respectively. Both genera are the only melectines to possess two submarginal cells in the forewing but are otherwise wholly dissimilar. Brachymelecta mucida (Cresson), a species known only from the male holotype collected in “Nevada”, is newly described and figured, including the first account of the hidden sterna and genitalia. Sinomelecta oreina Baker is similarly described and figured based on the holotype male and paratype female, apparently collected from the eastern Tibetan Plateau. Both genera are valid and from the available data do not appear to represent merely autapomorphic forms of Melecta Latreille. Indeed, the terminalia of Sinomelecta oreina are in some respects more similar to those of species of Thyreus Panzer.     

d-Ribulose-5-Phosphate 3-Epimerase: Cloning and Heterologous Expression of the Spinach Gene, and Purification and Characterization of the Recombinant Enzyme

We have achieved, to our knowledge, the first high-level heterologous expression of the gene encoding d-ribulose-5-phosphate 3-epimerase from any source, thereby permitting isolation and characterization of the epimerase as found in photosynthetic organisms. The extremely labile recombinant spinach (Spinacia oleracea L.) enzyme was stabilized by dl-α-glycerophosphate or ethanol and destabilized by d-ribulose-5-phosphate or 2-mercaptoethanol. Despite this lability, the unprecedentedly high specific activity of the purified material indicates that the structural integrity of the enzyme is maintained throughout isolation. Ethylenediaminetetraacetate and divalent metal cations did not affect epimerase activity, thereby excluding a requirement for the latter in catalysis. As deduced from the sequence of the cloned spinach gene and the electrophoretic mobility under denaturing conditions of the purified recombinant enzyme, its 25-kD subunit size was about the same as that of the corresponding epimerases of yeast and mammals. However, in contrast to these other species, the recombinant spinach enzyme was octameric rather than dimeric, as assessed by gel filtration and polyacrylamide gel electrophoresis under nondenaturing conditions. Western-blot analyses with antibodies to the purified recombinant enzyme confirmed that the epimerase extracted from spinach leaves is also octameric.As a participant in the oxidative pentose phosphate pathway, Ru5P epimerase (EC 5.1.3.1), which catalyzes the interconversion of Ru5P and Xu5P, is widely distributed throughout nature. Beyond its catabolic role, the epimerase is also vital anabolically to photosynthetic organisms in the regenerative phase of the reductive pentose phosphate pathway (the Calvin cycle). In this capacity, Ru5P epimerase directs Xu5P, formed in two distinct transketolase reactions of the cycle, to Ru5P. Phosphorylation of the latter regenerates d-ribulose-1,5-bisphosphate, the substrate for net CO₂ fixation. Because both the oxidative and reductive pentose phosphate pathways coexist in chloroplasts (Schnarrenberger et al., 1995), Ru5P epimerase and R5P isomerase facilitate partitioning of pentose phosphates between the two pathways, as dictated by the metabolic needs and redox status of the cell.Scant structural and mechanistic information about Ru5P epimerase is available despite its inherent importance and dual metabolic roles. This neglect may in part reflect the low natural abundance of the enzyme. For example, achievement of electrophoretic homogeneity required a 2000-fold purification from yeast (Bär et al., 1996) and spinach (Spinacia oleracea L.) chloroplasts (Teige et al., 1998) and 9000-fold purification from beef liver (Terada et al., 1985). Although low overall recoveries (<10%) further limited the availability of pure material, molecular sieving and denaturing electrophoresis established that the epimerases from mammals (Wood, 1979; Karmali et al., 1983; Terada et al., 1985) and yeast (Bär et al., 1996) are homodimers of approximately 23-kD subunits, whereas the enzyme from spinach chloroplasts may be an octamer of 23-kD subunits (Teige et al., 1998). DNA-deduced amino acid sequences of Ru5P epimerases from both photosynthetic and nonphotosynthetic sources, which confirm this estimated subunit size, show greater than 50% similarities among the most evolutionarily distant species examined (Kusian et al., 1992; Blattner et al., 1993; Falcone and Tabita, 1993; Lyngstadaas et al., 1995; Nowitzki et al., 1995; Teige et al., 1995).Although Ru5P epimerase has very recently been purified from a photosynthetic organism (spinach) for the first time (Teige et al., 1998), the low recovery (100 μg from 3.8 g of soluble chloroplast protein, representing an overall yield of 5%) imposes severe constraints on the directions of future experiments. Furthermore, despite successful cloning of cDNA fragments encoding Ru5P epimerase of several photosynthetic organisms (Kusian et al., 1992; Nowitzki et al., 1995; Teige et al., 1995), to our knowledge high-level heterologous expression and purification of enzymically active recombinant enzyme have not been achieved. Because of our interest in the regulation of photosynthetic carbon assimilation and the requisite need for ample supplies of the participant enzymes for use in mechanistic studies, we have attempted to optimize the heterologous expression of the spinach gene for Ru5P epimerase. In this paper we report cDNA clones that encode the mature chloroplastic enzyme or its cytoplasmic precursor. We also describe an efficient isolation procedure for the mature spinach enzyme synthesized in Escherichia coli and some of the properties of the purified enzyme. Contrasting features of the plant Ru5P epimerase, relative to the animal and yeast counterparts, include an octameric rather than a dimeric structure (also see Teige et al., 1998) and striking instability under routine laboratory conditions.     

Nematode Population Attrition and Histopathology of Heterodera glycines-Soybean Associations

Selected populations of soybean cyst nematodes were inoculated to roots of compatible and incompatible soybeans. Rates of penetration of infective juveniles of nematode populations selected on PI 209332, PI 89772, and Pickett 71 were equivalent on compatible and incompatible soybean roots. The first two populations averaged about 10% and the last about 5% penetration in 24-hour inoculations of young seedlings. About 14% of those juveniles that entered roots in compatible combinations developed into maturing females, compared with only about 1% in incompatible combinations. Several aberrations from the pattern of syncytial development associated with mature females in compatible hosts were apparent. A rapid necrotic response occurred in both kinds of hosts but was more frequent in incompatible associations. Delayed necrosis and small syncytia were present in some combinations. Those few females that developed in incompatible soybeans were associated with a characteristic syncytium different from the kind seen in roots of compatible hosts.     

Plant-parasitic Nematode Acetylcholinesterase Inhibition by Carbamate and Organophosphate Nematicides

The sensitivity of acetylcholinesterases (ACHE) isolated from the plant-parasitic nematodes Meloidogyne arenaria, M. incognita, and Heterodera glycines and the free-living nematode Caenorhabditis elegans to carbamate and organophosphate nematicides was examined. The AChE from plant-parasitic nematode species were more sensitive to carbamate inhibitors than was AChE from C. elegans, but response to the organophosphates was approximately equivalent. The sulfur-containing phosphate nematicides were poor inhibitors of nematode acetylcholinesterase, but treatment with an oxidizing agent greatly improved inhibition. Behavioral bioassays with living nematodes revealed a poor relationship between enzyme inhibition and expression of symptoms in live nematodes.     

Photoperiod Control of Gibberellin Levels and Flowering in Sorghum

Regulation of rhythmic peaks in levels of endogenous gibberellins (GAs) by photoperiod was studied in the short-day monocot sorghum (Sorghum bicolor [L.] Moench). Comparisons were made between three maturity (Ma) genotypes: 58M (Ma₁Ma₁, Ma₂Ma₂, phyB-1phyB-1, and Ma₄Ma₄ [a phytochrome B null mutant]); 90M (Ma₁Ma₁, Ma₂Ma₂, phyB-2phyB-2, and Ma₄Ma₄); and 100M (Ma₁Ma₁, Ma₂Ma₂, PHYBPHYB, and Ma₄Ma₄). Plants were grown for 14 d under 10-, 14-, 16-, 18-, and 20-h photoperiods, and GA levels were assayed by gas chromatography-mass spectrometry every 3 h for 24 h. Under inductive 10-h photoperiods, the peak of GA₂₀ and GA₁ levels in 90M and 100M was shifted from midday, observed earlier with 12-h photoperiods, to an early morning peak, and flowering was hastened. In addition, the early morning peaks in levels of GA₂₀ and GA₁ in 58M under conditions allowing early flowering (10-, 12-, and 14-h photoperiods) were shifted to midday by noninductive (18- and 20-h) photoperiods, and flowering was delayed. These results are consistent with the possibility that the diurnal rhythm of GA levels plays a role in floral initiation and may be one way by which the absence of phytochrome B causes early flowering in 58M under most photoperiods.     

Ecdysteroids in Axenically Propagated Caenorhabditis elegans and Culture Medium

Ecdysteroids (insect molting hormones) from Caenorhabditis elegans were chromatographically purified and quantified by radioimmunoassay. Nematodes from semidefined medium contained the immunoreactive equivalent of 460 pg ecdysone per gram dry weight. Culture medium, however, contained the immunoreactive equivalent of 68 times the quantity within the nematodes. In a defined medium lacking immunoreactivity, C. elegans contained 520 pg ecdysone equivalents per gram dry weight but reproduced slowly. Reproduction of C. elegans in defined medium was enhanced by formulation in agar. Propagation of C. elegans in either agar-based or aqueous defined medium supplemented with [¹⁴C]cholesterol of high specific activity failed to result in production of radiolabeled free ecdysteroids or polar or apolar ecdysteroid conjugates. Failure to demonstrate ecdysteroid biosynthesis in C. elegans raises questions about the ecdysteroids identified previously in nematodes being products of endogenous biosynthesis, a necessary condition for these compounds to be nematode hormones.     

Influence of Urea,Hydroxyurea, and Thiourea on Meloidogyne javanica and Infected Excised Tomato Roots in Culture

Urea (U), hydroxyurea (HU), and thiourea (TU), in various concentrations, were added to chemically defined plant tissue culture medium on which Meloidogyne javanica was reared on excised tomato roots. Concentrations as low as 3 ppm HU or 12 ppm TU inhibited nematode maturation by 70-90% 4 weeks after inoculation, and the coenocytes in the parasitized tissue were poorly developed. Gall weight was also inhibited by 50% in cultures treated with 3 and 6 ppm HU. However, exposing juveniles of M. javanica and Tylenchulus semipenetrans or juveniles and adults of Pratylenchus thornei to increasing concentrations of HU or TU, up to 100 ppm, was not lethal. These two urea derivatives still inhibited nematode maturation when the infected region of the root was not in direct contact with the chemicals. Therefore, we suggest that these urea derivatives inhibit nematode development by affecting the plant metabolism essential to coenocyte formation, an occurrence similar to the hypersensitive reaction in a naturally resistant plant.     

The Mechanism of Eukaryotic Translation Initiation: New Insights and Challenges

Translation initiation in eukaryotes is a highly regulated and complex stage of gene expression. It requires the action of at least 12 initiation factors, many of which are known to be the targets of regulatory pathways. Here we review our current understanding of the molecular mechanics of eukaryotic translation initiation, focusing on recent breakthroughs from in vitro and in vivo studies. We also identify important unanswered questions that will require new ideas and techniques to solve.This work aims to present the current state of our knowledge of the molecular mechanics of translation initiation in eukaryotes. We focus on advances that have taken place over the last few years and, because of space limitations, assume readers will be able to find references to the foundational literature for the field (published before 2000) in the more recent works that are cited here. As always, we apologize for not having the space to cite many important works. Please view this as merely an introduction to the field rather than a complete summary.     

The Chromatin-Remodeling Factor PICKLE Integrates Brassinosteroid and Gibberellin Signaling during Skotomorphogenic Growth in Arabidopsis

Plant cell elongation is controlled by endogenous hormones, including brassinosteroid (BR) and gibberellin (GA), and by environmental factors, such as light/darkness. The molecular mechanisms underlying the convergence of these signals that govern cell growth remain largely unknown. We previously showed that the chromatin-remodeling factor PICKLE/ENHANCED PHOTOMORPHOGENIC1 (PKL/EPP1) represses photomorphogenesis in Arabidopsis thaliana. Here, we demonstrated that PKL physically interacted with PHYTOCHROME-INTERACTING FACTOR3 (PIF3) and BRASSINAZOLE-RESISTANT1 (BZR1), key components of the light and BR signaling pathways, respectively. Also, this interaction promoted the association of PKL with cell elongation–related genes. We found that PKL, PIF3, and BZR1 coregulate skotomorphogenesis by repressing the trimethylation of histone H3 Lys-27 (H3K27me3) on target promoters. Moreover, DELLA proteins interacted with PKL and attenuated its binding ability. Strikingly, brassinolide and GA₃ inhibited H3K27me3 modification of histones associated with cell elongation–related loci in a BZR1- and DELLA-mediated manner, respectively. Our findings reveal that the PKL chromatin-remodeling factor acts as a critical node that integrates light/darkness, BR, and GA signals to epigenetically regulate plant growth and development. This work also provides a molecular framework by which hormone signals regulate histone modification in concert with light/dark environmental cues.     

Pinewood Nematode Species Complex: Interbreeding Potential and Chromosome Number

Interbreeding potential, chromosome number, and host range were compared among several isolates and species of Bursaphelenchus from diverse geographic areas. Some isolates from North America, Japan, and France had a wide-ranging interbreeding potential, whereas others were restricted in their potential to hybridize with other isolates. Although interbreeding occurred in the laboratory between some "M" and "R" forms of B. xylophilus, interbreeding of B. xylophilus and B. mucronatus was rare. The hybrids had the pathogenicity of the parent with the broader host range. This fact suggests that virulence may be inherited as a dominant character or that increased virulence may have resulted from differences in hybrid vigor. The haploid chromosome number of the different isolates separated the isolates into three groups and distinguished B. xylophilus from B. mucronatus. The findings suggest that the pinewood nematode species complex consists of sibling species that have evolved by reproductive isolation, that the French isolate is a new species, and that B. xylophilus and B. mucronatus have evolved from a common ancestor.     

The Boron Requirement and Cell Wall Properties of Growing and Stationary Suspension-Cultured Chenopodium album L. Cells

Suspension-cultured Chenopodium album L. cells are capable of continuous, long-term growth on a boron-deficient medium. Compared with cultures grown with boron, these cultures contained more enlarged and detached cells, had increased turbidity due to the rupture of a small number of cells, and contained cells with an increased cell wall pore size. These characteristics were reversed by the addition of boric acid (≥7 μm) to the boron-deficient cells. C. album cells grown in the presence of 100 μm boric acid entered the stationary phase when they were not subcultured, and remained viable for at least 3 weeks. The transition from the growth phase to the stationary phase was accompanied by a decrease in the wall pore size. Cells grown without boric acid or with 7 μm boric acid were not able to reduce their wall pore size at the transition to the stationary phase. These cells could not be kept viable in the stationary phase, because they continued to expand and died as a result of wall rupture. The addition of 100 μm boric acid prevented wall rupture and the wall pore size was reduced to normal values. We conclude that boron is required to maintain the normal pore structure of the wall matrix and to mechanically stabilize the wall at growth termination.The ultrastructure and physical properties of plant cell walls are known to be affected by boron deficiency (Kouchi and Kumazawa, 1976; Hirsch and Torrey, 1980; Fischer and Hecht-Buchholz, 1985; Matoh et al., 1992; Hu and Brown, 1994; Findeklee and Goldbach, 1996). Moreover, boron is predominantly localized in the cell wall when plants are grown with suboptimal boron (Loomis and Durst, 1991; Matoh et al., 1992; Hu and Brown, 1994; Hu et al., 1996). In radish, >80% of the cell wall boron is present in the pectic polysaccharide RG-II (Matoh et al., 1993; Kobayashi et al., 1996), which is now known to exist as a dimer that is cross-linked by a borate ester between two apiosyl residues (Kobayashi et al., 1996; O''Neill et al., 1996). Dimeric RG-II is unusually stable at low pH and is present in a large number of plant species (Ishii and Matsunaga, 1996; Kobayashi et al., 1996, 1997; Matoh et al., 1996; O''Neill et al., 1996; Pellerin et al., 1996; Kaneko et al., 1997). The widespread occurrence and conserved structure of RG-II (Darvill et al., 1978; O''Neill et al., 1990) have led to the suggestion that borate ester cross-linked RG-II is required for the development of a normal cell wall (O''Neill et al., 1996; Matoh, 1997).One approach for determining the function of boron in plant cell walls is to compare the responses to boron deficiency of growing plant cells that are dividing and synthesizing primary cell walls with those of growth-limited plant cells in which the synthesis of primary cell walls is negligible. Suspension-cultured cells are well suited for this purpose because they may be reversibly transferred from a growth phase to a stationary phase. Continuous cell growth phase is maintained by frequent transfer of the cells into new growth medium (King, 1981; Kandarakov et al., 1994), whereas a stationary cell population is obtained by feeding the cells with Suc and by not subculturing them. Cells in the stationary phase are characterized by mechanically stabilized primary walls and reduced biosynthetic activity. Here we describe the responses of suspension-cultured Chenopodium album L. cells in the growth and stationary phases to boron deficiency. These cells have a high specific-growth rate, no significant lag phase, and reproducible changes in their wall pore size during the transition from the growth phase to the stationary phase (Titel et al., 1997).