Uses,traditional management,perception of variation and preferences in ackee (Blighia sapida K.D. Koenig) fruit traits in Benin: implications for domestication and conservation

Background

Changing lifestyles have recently caused a severe reduction of the gathering of wild food plants. Knowledge about wild food plants and the local environment becomes lost when plants are no longer gathered. In Central Europe popular scientific publications have tried to counter this trend. However, detailed and systematic scientific investigations in distinct regions are needed to understand and preserve wild food uses. This study aims to contribute to these investigations.

Methods

Research was conducted in the hill country east of Graz, Styria, in Austria. Fifteen farmers, most using organic methods, were interviewed in two distinct field research periods between July and November 2008. Data gathering was realized through freelisting and subsequent semi-structured interviews. The culinary use value (CUV) was developed to quantify the culinary importance of plant species. Hierarchical cluster analysis was performed on gathering and use variables to identify culture-specific logical entities of plants. The study presented was conducted within the framework of the master's thesis about wild plant gathering of the first author. Solely data on gathered wild food species is presented here.

Results

Thirty-nine wild food plant and mushroom species were identified as being gathered, whereas 11 species were mentioned by at least 40 percent of the respondents. Fruits and mushrooms are listed frequently, while wild leafy vegetables are gathered rarely. Wild foods are mainly eaten boiled, fried or raw. Three main clusters of wild gathered food species were identified: leaves (used in salads and soups), mushrooms (used in diverse ways) and fruits (eaten raw, with milk (products) or as a jam).

Conclusions

Knowledge about gathering and use of some wild food species is common among farmers in the hill country east of Graz. However, most uses are known by few farmers only. The CUV facilitates the evaluation of the culinary importance of species and makes comparisons between regions and over time possible. The classification following gathering and use variables can be used to better understand how people classify the elements of their environment. The findings of this study add to discussions about food heritage, popularized by organizations like Slow Food, and bear significant potential for organic farmers.     

Preparation of DNA suitable for PCR amplification from fresh or fixed single dinoflagellate cells

A method is described to prepare total DNA from single cells of dinoflagellates, which can be used for PCR amplification. As model organisms, we used a stock strain of Alexandrium catenella and cells of Dinophysis acuminata harvested from the Atlantic Ocean. Fresh grown cells or cells maintained in different preservatives were tested as sources for DNA preparation. The method used to prepare DNA combines physicochemical and enzymatic procedures on cells embedded in agarose plugs or beads. The agarose pieces containing the DNA were used to perform PCR amplification of a fragment of DNA containing a 5.8S rRNA gene and the flanking internal transcribed spacers (ITS1 and ITS2).     

Voltammetric study of conductive planar assemblies of Geobacter nanowire pilins unmasks their ability to bind and mineralize divalent cobalt

Journal of Industrial Microbiology & Biotechnology - Geobacter bacteria assemble a helical peptide of the Type IVa pilin subclass as conductive pili decorated with metal binding and reduction...     

Evolution of eutherian cytochrome c oxidase subunit II: heterogeneous rates of protein evolution and altered interaction with cytochrome c

Cytochrome c oxidase subunit II (COII), encoded by the mitochondrial genome, exhibits one of the most heterogeneous rates of amino acid replacement among placental mammals. Moreover, it has been demonstrated that cytochrome c oxidase has undergone a structural change in higher primates which has altered its physical interaction with cytochrome c. We collected a large data set of COII sequences from several orders of mammals with emphasis on primates, rodents, and artiodactyls. Using phylogenetic hypotheses based on data independent of the COII gene, we demonstrated that an increased number of amino acid replacements are concentrated among higher primates. Incorporating approximate divergence dates derived from the fossil record, we find that most of the change occurred independently along the New World monkey lineage and in a rapid burst before apes and Old World monkeys diverged. There is some evidence that Old World monkeys have undergone a faster rate of nonsynonymous substitution than have apes. Rates of substitution at four-fold degenerate sites in primates are relatively homogeneous, indicating that the rate heterogeneity is restricted to nondegenerate sites. Excluding the rate acceleration mentioned above, primates, rodents, and artiodactyls have remarkably similar nonsynonymous replacement rates. A different pattern is observed for transversions at four-fold degenerate sites, for which rodents exhibit a higher rate of replacement than do primates and artiodactyls. Finally, we hypothesize specific amino acid replacements which may account for much of the structural difference in cytochrome c oxidase between higher primates and other mammals.      

Mutational analysis of methionine adenosyltransferase from Leishmania donovani.

The methionine adenosyltransferase (MAT; EC 2.5.1.6) mediated synthesis of S-adenosylmethionine (AdoMet) is a two-step process, consisting of the formation of AdoMet and the subsequent cleavage of the tripolyphosphate (PPPi) molecule, a reaction induced, in turn, by AdoMet. The fact that the two activities--AdoMet synthesis and tripolyphosphate hydrolysis--can be measured separately is particularly useful when the site-directed mutagenesis approach is used to determine the functional role of the amino acid residues involved in each. This report describes the mutational analysis of the amino acids involved in both the ATP and L-methionine binding sites of Leishmania donovani MAT (GenBank accession number AF179714) the aetiological agent of visceral leishmaniasis. Site-directed mutagenesis was used to substitute neutral residues for the basic amino acid (Lys168, Lys256, Lys276, Lys280 and His17), acidic residues (Asp19, Asp121, Asp166, Asp249, Asp277 and Asp288) and Phe241 involved in AdoMet synthesis and PPPi hydrolysis. With the exception of D116N, none of these mutants was able to synthesize AdoMet at a significant rate, although H17A, H17N, K256A, K280A, D19N, D121N, D166N, D249N and D282N showed measurable tripolyphosphatase activity. Finally, the C-terminus domain of L. donovani MAT was truncated at three points (F382Stop, D375Stop, F368Stop), deleting a 3(10) one-turn helix motif in all three cases. Whilst none of the truncated proteins conserved MAT activity, they were able to hydrolyse PPPi, albeit at a lower rate than the wild-type enzyme. A fourth protein with an internal deletion (E376DeltaF382) in the C-terminal domain conserved high tripolyphosphatase activity, which was not, however, induced by 50 microM AdoMet.     

Identification of a gene in Leishmania infantum encoding a protein that contains a SP-RING/MIZ zinc finger domain

The SP-RING or Miz zinc finger domain that is related to the classical RING-finger motif, defines a class of proteins that can act as E3-like factors in the pathway of small ubiquitin-related modifier (SUMO) conjugation. This family includes the mammalian protein inhibitor of activated STAT (PIAS) proteins and related proteins from lower eukaryotes. Here we report the existence of a gene in Leishmania infantum, present as two identical copies placed upstream of each MAT2 gene copy, and transcribed as a single approximately 2.2 kb mRNA both in the logarithmic and stationary phases of the promastigote stage. This gene encodes a 47 kDa protein that has been named LORIEN. LORIEN is circumscribed to the cell periphery and it is antigenic during L. infantum infection of dogs and hamsters. Strikingly, this novel protein contains a highly conserved SP-RING/Miz zinc finger domain, raising the possibility that a SUMO or ubiquitin-like system may exist in this microorganism.     

SMALL CELL AND INTERMEDIATE CELL FORMATION IN SPECIES OF DINOPHYSIS (DINOPHYCEAE, DINOPHYSIALES)

Observations of two distinct size classes with similar shape in natural populations of Dinophysis Ehrenberg were first reported by Jorgensen in 1923 and intermediate forms exhibiting a continuum between the typical vegetative cell and a putative small cell by Wood in 1954. Focused attention on Dinophysis spp. associated with diarrhetic shellfish intoxications in the last decade has provided new examples of small cells in the genus, sometimes with contours dissimilar from the corresponding vegetative cells; dimorphic individuals; and large/small cell couplets. This work was based on in situ observations during intensive sampling for cell cycle studies of Dinophysis acuminata Claparéde et Lachmann, Dinophysis acuta Ehrenberg, Dinophysis caudata Saville-Kent, and Dinophysis tripos Gourret; on laboratory incubations of D. acuminata; and on a thorough search of documented information on morphological variability of Dinophysis spp. During in situ division, most dividing cells exhibit a normal longitudinal fission, but some (1%–10%) undergo a “depauperating” fission, leading to pairs of dimorphic cells with dissimilar moieties. After separation and sulcal list regeneration, these dimorphic cells become D. skagii Paulsen, D. dens Pavillard, D. diegensis Kofoid, and D. diegensis Kofoid var. curvata-like individuals, which can also be observed forming couplets D. acuminata/D. skagii, D. acuta/D. dens, and D. caudata/D. diegensis attached by their ventral margins. Small cells can grow again to large size, as shown in laboratory incubations of D. acuminata, thus partly explaining observations of thecal intercalary bands, and intermediate forms. The sexual nature of the small cells will not be unequivocally demonstrated until controlled germination of the alleged cyst forms is achieved, and some intermediate forms may correspond to undescribed stages after cyst germination. These observations suggest common patterns in the life cycle of Dinophysis spp. Intraspecific morphological variability of Dinophysis spp. in a given geographic area can largely be attributed to small cell formation, as a response to changing environmental conditions, and may be a part of the sexual cycle of these species. Small cells seem to be able to enlarge, leading to intermediate cell and further vegetative cell formation as part of a three-looped life history pattern in Dinophysis.     

Evidence that biosynthesis of phosphatidylethanolamine, phosphatidylcholine, and triacylglycerol occurs on the cytoplasmic side of microsomal vesicles

Experiments were performed to localize the hepatic microsomal enzymes of phosphatidylcholine, phosphatidylethanolamine, and triacylglycerol biosynthesis to the cytoplasmic or lumenal surface of microsomal vesicles. Greater than 90 percent of the activities of fatty acid-CoA ligase (EC 6.2.1.3), sn-glycerol 3-phosphate acyltransferase (EC 2.3.1.15), lysophosphatidic acid acyltransferase, diacylglycerol acyltransferase (EC 2.3.1.20), diacylglycerol cholinephosphotransferase (EC 2.7.8.2), and diacylglycerol ethanolaminephosphotransferase (EC 2.7.8.1) was inactivated by proteolysis of intact microsomal vesicles. The phosphatidic acid phosphatase (EC 3.1.3.4) was not inactivated by any of the protease tested. Under conditions employed, <5 percent of the luminal mannose-6-phosphatase (EC 3.1.3.9) activity was lost. After microsomal integrity was disrupted with detergents, protease treatment resulted in a loss of >74 percent of the mannose-6-phosphatase activity. The latency of the mannose-6-phosphatase activity was not affected by protease treatment. Mannose-6-phosphatase latency was not decreased by the presence of the assay components of several of the lipid biosynthetic activities, indicating that those components did not disrupt the microsomal vesicles. None of the lipid biosynthetic activities appeared latent. The presence of a protease-sensitive component of these biosynthetic activities on the cytoplasmic surface of microsomal vesicles, and the absence of latency for any of these biosynthetic activities suggest that the biosynthesis of phosphatidylcholine, phosphatidylethanolamine, and triacylglycerol occurs asymmetrically on the cytoplasmic surface of the endoplasmic reticulum. The location of biosynthetic activities within the transverse plane of the endoplasmic reticulum is of particular interest for enzymes whose products may be either secreted or retained within the cell. Phosphatidylcholine, phosphatidylethanolamine, and triacylglycerol account for the vast majority of hepatic glycerolipid biosynthesis. The phospholipids are utilized for hepatic membrane biogenesis and for the formation of lipoproteins, and the triacylglycerols are incorporated into lipoproteins or accumulate within the hepatocyte in certain disease states (14). The enzymes responsible for the biosynthesis of these glycerolipids (Scheme I) from fatty acids and glycerol-3P have all been localized to the microsomal subcellular fraction (12, 16, 29, 30). Microsomes are derived from the endoplasmic reticulum and are sealed vesicles which maintain proper sidedness. (11, 22). The external surface of these vesicles corresponds to the cytoplasmic surface of the endoplasmic reticulum. Macromolecules destined for secretion must pass into the lumen of the endoplasmic reticulum (5, 23). Uncharged molecules of up to approximately 600 daltons are able to enter the lumen of rat liver microsomes, but macromolecules and charged molecules of low molecular weight do not cross the vesicle membrane (10, 11). Because proteases neither cross the microsomal membrane nor destroy the permeability barrier of the microsomal vesicles, only the enzymes and proteins located on the cytoplasmic surface of microsomal vesicles are susceptible to proteolysis unless membrane integrity is disrupted (10, 11). By use of this approach, several enzymes and proteins have been localized in the transverse plane of microsomal membranes (11). With the possible exception of cytochrome P 450, all of the enzymes and proteins investigated were localized asymmetrically by the proteolysis technique (11). By studies of this type, as well as by product localization, glucose-6-phosphate (EC 3.1.3.9) has been localized to the luminal surface of microsomal vesicles (11) and of the endoplasmic reticulum (18, 19). All microsomal vesicles contain glucose-6-phosphatase (18, 19) which can effectively utilize mannose-6-P as a substrate, provided the permeability barrier of the vesicles has been disrupted to allow the substrate access to the active site located on the lumenal surface (4). An exact correspondence between mannose- 6-phosphate activity and membrane permeability to EDTA has been established (4). The latency of mannose-6-phosphatase activity provides a quantitative index of microsomal integrity (4.) Few of the microsomal enzymes in the synthesis of phosphatidylcholine, phosphatidylethanolamine, and triacylglycerol have been solubilized and/or purified, and little is known about the topography of these enzymes in the transverse or lateral planes of the endoplasmic reticulum. An asymmetric location of these biosynthetic enzymes on the cytoplasmic or lumenal surface of microsomal vesicles may provide a mechanism for regulation of the glycerolipids to be retained or secreted by the cell, and for the biogenesis of asymmetric phospholipid bilayers. In this paper, we report investigations on the localization of all seven microsomal enzymes (Scheme I) in the biosynthesis of triacylglycerol, phosphatidylcholine, and phosphatidylethanolamine, using the protease technique with mannose-6-phosphatase serving as luminal control activity. The latency of these lipid biosynthetic enzymes was also investigated, using the latency of mannose-6-phosphatase as an index of microsomal integrity.     

Adding Content to Contacts: Measurement of High Quality Contacts for Maternal and Newborn Health in Ethiopia,North East Nigeria,and Uttar Pradesh,India

BackgroundFamilies in high mortality settings need regular contact with high quality services, but existing population-based measurements of contacts do not reflect quality. To address this, in 2012, we designed linked household and frontline worker surveys for Gombe State, Nigeria, Ethiopia, and Uttar Pradesh, India. Using reported frequency and content of contacts, we present a method for estimating the population level coverage of high quality contacts.ConclusionsMeasuring content of care to reflect the quality of contacts can reveal missed opportunities to deliver best possible health care.