Partial isolation from intact cells of a cell surface-exposed lysophosphatidylinositol-phospholipase C

A novel cell surface phosphoinositide-cleaving phospholipase C (ecto-PLC) activity was isolated from cultured cells by exploiting its presumed external exposure. Biotinylation of intact cells followed by solubilization of the biotinylated proteins from a membrane fraction and recovery onto immobilized-avidin beads, allowed assay of this cell surface enzyme activity apart from the background of the substantial family of intracellular PLCs. Several cell lines of differing ecto-PLC expression were examined as well as cells stably transfected to overexpress the glycosylphosphatidylinositol (GPI)-anchored protein human placental alkaline phosphatase (PLAP) as a cell surface enzyme marker. The resulting bead preparations from ecto-PLC positive cells possessed calcium-dependent PLC activity with preference for lysophosphatidylinositol (lysoPI) rather than phosphatidylinositol (PI). The function of ecto-PLC of intact cells evidently is not to release GPI-anchored proteins at the cell surface, as no detectable Ca²⁺-dependent release of overexpressed PLAP from ecto-PLC-positive cells was observed. To investigate the cell surface linkage of the ecto-PLC itself, intact cells were treated with bacterial PI-PLC to cleave simple GPI anchors, but no decrease in ecto-PLC activity was observed. High ionic strength washes of biotinylated membranes prior to the generation of bead preparations did not substantially reduce the lysoPI-PLC activity. The results verify that the ecto-PLC is truly cell surface-exposed, and unlike other members of the PLC family that are thought to be peripheral membrane proteins, this novel lysoPI-PLC is most likely a true membrane protein. J. Cell. Biochem. 65:550–564. © 1997 Wiley-Liss Inc.     

Use of the additive main effects and multiplicative interaction model in QTL mapping for adaptation in barley

The additive main effects and multiplicative interaction (AMMI) model has emerged as a powerful analytical tool for genotype x environment studies. The objective of the present study was to assess its value in quantitative trait locus (QTL) mapping. This was done through the analysis of a large two-way table of genotype-by-environment data of barley (Hordeum vulgare L.) grain yields, where the genotypes constituted a genetic population suitable for mapping studies. Grain yield data of 150 doubled haploid lines derived from the Steptoe x Morex cross, and the two parental lines, were taken by the North American Barley Genome Mapping Project (NABGMP) at 16 environments throughout the barley production areas of the USA and Canada. Four regions of the genome were responsible for most of the differential genotypic expression across environments. They accounted for approximately 50% of the genotypic main effect and 30% of the genotype x environment interaction (GE) sums of squares. The magnitude and sign of AMMI scores for genotypes and sites facilitate inferences about specific interactions. The parallel use of classification (cluster analysis of environments) and ordination (principal component analysis of GE matrix) techniques allowed most of the variation present in the genotype x environment matrix to be summarized in just a few dimensions, specifically four QTLs showing differential adaptation to four clusters of environments. Thus, AMMI genotypic scores, when the genotypes constituted a population suitable for QTL mapping, could provide an adequate way of resolving the magnitude and nature of QTL x environment interactions.Ignacio Romagosa was on sabbatical leave from the University of Lleida and the Institut de Recerca i Tecnologia Agroalimentàries, Lleida, Spain, when this study was conducted     

Effects of Radio-Collars are not Contingent on Socioecological Conditions in Degus

Species-specific research on free-ranging mammals reveals a diversity of effects of radio-collars on behavior, body condition, and fitness. Although these studies indicate rather limited direct effects, radio-collars may cause effects influenced by socio-ecological conditions. Using a 7-year study on a natural population of group-living degus (Octodon degus), we tested the hypothesis that ecological (food availability, burrow density) and social (group size, group male-to-female ratio) conditions modulate effects of radio-collars on body condition (e.g., body mass, ecto- and endoparasite loads, fecal cortisol metabolites) and direct fitness (litter size, adult survival). We determined the effect of radio-collar use on degus by contrasting the presence or absence of radio-collars, quantifying the effects of the number of days carrying a radio-collar, and the relative mass of radio-collars worn by degus in central Chile between 2009 and 2015. Radio-collar use was not associated with direct effects on litter size, adult survival, or with body mass and fecal cortisol metabolites but was linked to low ecto- and endoparasite loads. These seemingly positive effects may reflect decreased mobility, or a research bias for radio-collaring larger, healthier individuals. There was no evidence that ecological and social conditions modulated radio-collar effects on degu body condition and direct fitness. These findings are consistent with evidence from other mammal studies that reported no appreciable detrimental direct or indirect effects of radio-collars. © 2021 The Wildlife Society.     

A REAPPRAISAL OF PORPHYRA AND BANGIA (BANGIOPHYCIDAE, RHODOPHYTA) IN THE NORTHEAST ATLANTIC BASED ON THE rbcL–rbcS INTERGENIC SPACER

Sequence data of the rbc L –rbc S noncoding intergenic spacer of the plastid genome for 47 specimens of Porphyra and Bangia from the northeast Atlantic reveal that they fall into 11 distinct sequences: P. purpurea, P. dioica (includes a sample of P. "ochotensis" from Helgoland), P. amplissima (includes P. thulaea and British records of P. "miniata" ), P. linearis, P. umbilicalis, P. "miniata", B. atropurpurea s.l. from Denmark and B. atropurpurea s.l. from Wales, P. drachii, P. leucosticta (includes a British record of P. "miniata var. abyssicola" ), and P. "insolita" (includes P. "yezoensis" from Helgoland). Of these, data obtained for P. purpurea , P. dioica, P. amplissima, P. linearis, P. umbilicalis, P. drachii, and P. leucosticta were based on type specimens or material compared with types. Comparison of sequence data for Porphyra spp. and Bangia atropurpurea s.l. (including B. fuscopurpurea, the type species of Bangia ) confirms that the species are congeneric. The data also confirm that the number of layers that make up the Porphyra thallus are not taxonomically significant. Comparison of sequence data for species from the northeast Atlantic with those for material of two species from the Pacific reveals that the species fall into two distinct groupings: an Atlantic group, containing P. purpurea, P. dioica, P. amplissima, P. linearis, P. umbilicalis, P. "miniata", and B. atropurpurea, and a Pacific group, containing P. "pseudolinearis", P. drachii, P. leucosticta, P. "yezoensis" (including a sample of P. "tenera" ), and P. "insolita" (including P. "yezoensis" from Helgoland). The possibility of alien species in the northeast Atlantic is discussed.     

Characterizing associations and SNP-environment interactions for GWAS-identified prostate cancer risk markers--results from BPC3

Genome-wide association studies (GWAS) have identified multiple single nucleotide polymorphisms (SNPs) associated with prostate cancer risk. However, whether these associations can be consistently replicated, vary with disease aggressiveness (tumor stage and grade) and/or interact with non-genetic potential risk factors or other SNPs is unknown. We therefore genotyped 39 SNPs from regions identified by several prostate cancer GWAS in 10,501 prostate cancer cases and 10,831 controls from the NCI Breast and Prostate Cancer Cohort Consortium (BPC3). We replicated 36 out of 39 SNPs (P-values ranging from 0.01 to 10⁻²⁸). Two SNPs located near KLK3 associated with PSA levels showed differential association with Gleason grade (rs2735839, P = 0.0001 and rs266849, P = 0.0004; case-only test), where the alleles associated with decreasing PSA levels were inversely associated with low-grade (as defined by Gleason grade <8) tumors but positively associated with high-grade tumors. No other SNP showed differential associations according to disease stage or grade. We observed no effect modification by SNP for association with age at diagnosis, family history of prostate cancer, diabetes, BMI, height, smoking or alcohol intake. Moreover, we found no evidence of pair-wise SNP-SNP interactions. While these SNPs represent new independent risk factors for prostate cancer, we saw little evidence for effect modification by other SNPs or by the environmental factors examined.     

Esterification reactions of lipase in reverse micelles

The activities of lipase from Candida cylindracea and Rhizopus delemar have been investigated in water/AOT/iso-octane reverse micellar media through the use of two esterification reactions: fatty acid-alcohol esterification and glyceride synthesis. Such media promotes the occurrence of these two lipase-catalyzed reactions due to its low water content. The effect of various parameters on the activity of lipase from C. cylindracea in reverse micelles was determined and compared to results where alternate media were employed. It was observed that the structure of the media, as dictated by the type and concentration of the substrates and products and by the water/AOT ratio, w(0), had a strong impact on enzyme activity. Strong deactivation of both typase types occurred in reverse micelles, especially in the absence of substrates and for w(0) values greater than 3.0. Glyceride synthesis was realized with lipase from R. delemar, but not with that from C. cylindracea; the temperature and concentration of substrates and water strongly dictated the reaction rate and the percent conversion.     

Social Tolerance in Wild Female Crested Macaques (Macaca nigra) in Tangkoko‐Batuangus Nature Reserve,Sulawesi, Indonesia

In primates, females typically drive the evolution of the social system and present a wide diversity of social structures. To understand this diversity, it is necessary to document the consistency and/or flexibility of female social structures across and within species, contexts, and environments. Macaques (Macaca sp.) are an ideal taxon for such comparative study, showing both consistency and variation in their social relations. Their social styles, constituting robust sets of social traits, can be classified in four grades, from despotic to tolerant. However, tolerant species are still understudied, especially in the wild. To foster our understanding of tolerant societies and to assess the validity of the concept of social style, we studied female crested macaques, Macaca nigra, under entirely natural conditions. We assessed their degree of social tolerance by analyzing the frequency, intensity, and distribution of agonistic and affiliative behaviors, their dominance gradient, their bared‐teeth display, and their level of conciliatory tendency. We also analyzed previously undocumented behavioral patterns in grade 4 macaques: reaction upon approach and distribution of affiliative behavior across partners. We compared the observed patterns to data from other populations of grade 4 macaques and from species of other grades. Overall, female crested macaques expressed a tolerant social style, with low intensity, frequently bidirectional, and reconciled conflicts. Dominance asymmetry was moderate, associated with an affiliative bared‐teeth display. Females greatly tolerated one another in close proximity. The observed patterns matched the profile of other tolerant macaques and were outside the range of patterns of more despotic species. This study is the first comprehensive analysis of females’ social behavior in a tolerant macaque species under natural conditions and as such, contributes to a better understanding of macaque societies. It also highlights the relevance of the social style concept in the assessment of the degree of tolerance/despotism in social systems. Am. J. Primatol. 75:361‐375, 2013. © 2013 Wiley Periodicals, Inc.