The role of helix 8 and of the cytosolic C-termini in the internalization and signal transduction of B(1) and B(2) bradykinin receptors

Determinants for desensitization and sequestration of G protein-coupled receptors often contain serine or threonine residues located in their C-termini. The sequence context, however, in which these residues have to appear, and the receptor specificity of these motifs are largely unknown. Mutagenesis studies with the B(2) bradykinin receptor (B(2)wt), stably expressed in HEK 293 cells, identified a sequence distal to N338 (NSMGTLRTSI, including I347 but not the basally phosphorylated S348) and in particular the TSI sequence therein, as a major determinant for rapid agonist-inducible internalization and the prevention of receptor hypersensitivity. Chimeras of the noninternalizing B(1) bradykinin receptor (B(1)wt) containing these B(2)wt sequences sequestered poorly, however, suggesting that additional motifs more proximal to N338 are required. In fact, further substitution of the B(1)wt C-terminus with corresponding B(2)wt regions either at C330(7.71) following putative helix 8 (B(1)CB(2)) or at the preceding Y312(7.53) in the NPXXY sequence (B(1)YB(2)) resulted in chimeras displaying rapid internalization. Intriguingly, however, exchange performed at K322(7.63) within putative helix 8 generated a slowly internalizing chimera (B(1)KB(2)). Detailed mutagenesis analysis generating additional chimeras identified the change of V323 in B(1)wt to serine (as in B(2)wt) as being responsible for this effect. The slowly internalizing chimera as well as a B(1)wt point-mutant V323S displayed significantly reduced inositol phosphate accumulation as compared to B(1)wt or the other chimeras. The slow internalization of B(1)KB(2) was also accompanied by a lack of agonist-induced phosphorylation, that in contrast was observed for B(1)YB(2) and B(1)CB(2), suggesting that putative helix 8 is either directly or indirectly (e.g. via G protein activation) involved in the interaction between the receptor and receptor kinases.     

Biotechnology software in the digital age: are you winning?

There is a digital revolution taking place and biotechnology companies are slow to adapt. Many pharmaceutical, biotechnology, and industrial bio-production companies believe that software must be developed and maintained in-house and that data are more secure on internal servers than on the cloud. In fact, most companies in this space continue to employ large IT and software teams and acquire computational infrastructure in the form of in-house servers. This is due to a fear of the cloud not sufficiently protecting in-house resources and the belief that their software is valuable IP. Over the next decade, the ability to quickly adapt to changing market conditions, with agile software teams, will quickly become a compelling competitive advantage. Biotechnology companies that do not adopt the new regime may lose on key business metrics such as return on invested capital, revenue, profitability, and eventually market share.     

Atypical virulence in a type III Toxoplasma gondii strain isolated in Japan

The virulence of a type III Toxoplasma gondii strain isolated in Japan and designated here as TgCatJpGi1/TaJ was examined in mice and micro minipigs in this study. Despite its type III genotype, oral or intraperitoneal inoculation of cysts from it resulted in severe virulence in C57BL/6J and BALB/c mice. In contrast, mice inoculated with a high dose of TgCatJpGi1/TaJ tachyzoites showed no obvious clinical signs of infection, and all of them survived for >21?days post-inoculation. Furthermore, no clinical signs of infection were seen when micro minipigs were inoculated with 900 cysts. Interestingly, our allelic type screening of the virulence-related rop5, rop16, rop17, and rop18 genes, as based on restriction fragment length polymorphism analysis (RFLP), revealed that the RFLP patterns for TgCatJpGi1/TaJ were identical to those from nonvirulent type III parasites. These results suggest that TgCatJpGi1/TaJ possesses an unknown virulence factor or factors.     

Selection for life‐history traits to maximize population growth in an invasive marine species

Species establishing outside their natural range, negatively impacting local ecosystems, are of increasing global concern. They often display life‐history features characteristic for r‐selected populations with fast growth and high reproduction rates to achieve positive population growth rates (r) in invaded habitats. Here, we demonstrate substantially earlier maturation at a 2 orders of magnitude lower body mass at first reproduction in invasive compared to native populations of the comb jelly Mnemiopsis leidyi. Empirical results are corroborated by a theoretical model for competing life‐history traits that predicts maturation at the smallest possible size to optimize r, while individual lifetime reproductive success (R₀), optimized in native populations, is near constant over a large range of intermediate maturation sizes. We suggest that high variability in reproductive tactics in native populations is an underappreciated determinant of invasiveness, acting as substrate upon which selection can act during the invasion process.     

Gonenkonkurrenz bei Oenothera unter dem Einflu\ eines gametophytisch wirksamen Gens in der ersten Koppelungsgruppe sowie ein Modell für die Untersuchung verzweigter Koppelungsgruppen

Zusammenfassung Die Nachkommenschaften aus Rückkreuzungen des Pollens von Bastarden mit den durch zwei Test-gene und den Translokationspunkt markierten Komplexen ^h hookeri und flavens zeigen Spaltungszahlen, die gesichert von der Mendel-Erwartung abweichen.Es wird ein gametophytisch wirksamer Genlocus in der ersten Koppelungsgruppe angenommen, dessen Allele ga ⁺ und ga ^- so auf die Entwicklung der Gametophyten (Pollen und Pollenschläuche) einwirken, daß den Gameten mit verschiedenen ga-Allelen eine unterschiedliche Befruchtungswahrscheinlichkeit zukommt, wenn sich die Pollenkörner auf einer heterozygoten Mutterpflanze entwickelt haben.Es werden in mehreren Stufen genetische Modelle entwickelt, bei denen die den Allelen ga ⁺ und ga ^- zugeordneten Befruchtungswahrscheinlichkeiten, die Austauschwahrscheinlichkeiten auf den untersuchten Genstrecken und die Verzweigung der Koppelungsgruppe durch einen Translokationspunkt berücksichtigt werden.Anhand dieser Modelle werden Voraussagen über die zu erwartenden Relationen zwischen den relativen Häufigkeiten der einzelnen Gametenklassen unter verschiedenen Voraussetzungen über die Anordnung der Genloci in der Koppelungsgruppe möglich.Durch die Formulierung des Modells kann es über den vorliegenden Fall hinaus allgemein sowohl für die Lokalisation von gametophytisch wirksamen Loci als auch für die Lokalisation beliebiger Genloci in verzweigten Koppelungsgruppen verwendet werden.Es erweist sich als unmöglich, aus Rückkreuzungsdaten den Grad der Benachteiligung von ga ^- gegenüber ga ⁺ zu bestimmen. Aus empirischen Daten kann vielmehr nur ein Schätzwert gewonnen werden, in den die Befruchtungswahrscheinlichkeiten und der Austausch auf der Strecke zwischen Ga und dem Testlocus eingehen.Durch Vergleich der empirischen Daten mit den Modellen wird versucht, die Anordnung der Testloci und des Locus Ga in bezug auf den Translokationspunkt zu bestimmen. Die meisten Befunde sprechen für eine verzweigte Koppelungsgruppe, in der die drei Loci s, de und Ga auf drei Armen der Translokationsfigur angeordnet sind. Eine lineare Anordnung in zwei Chromosomenarmen kann jedoch nicht mit Sicherheit ausgeschlossen werden.Die Austauschwerte für die untersuchten Genstrekken zeigen eine erhebliche Variabilität zwischen Bastarden und zwischen verschiedenen Nachkommenschaften eines Bastards.

---

Competition between the male gametophytes in Oenothera under the influence of a gametophytic gene in the first linkage group and a model for investigations of branched genetic maps

Summary The progeny ratios obtained in male backcrosses of Oenothera hybrids to the complexes hookeri and flavens (identified by two marker genes and by the translocation break) showed statistically significant differences from Mendelian expectations.Presence of a gametophytic gene in the first linkage group is proposed. Its alleles ga ⁺ and ga ^- are thought to control development of the male gametophytes in such a manner that pollen grains carrying the different ga alleles and developing in a heterozygous female plant, will fertilize the female gamete with different average frequencies.A series of genetic models is developed, all taking into account the relative probability of the alleles ga ⁺ and ga ^- to function in fertilization, the probability of crossing over between the gene loci studied, and the branched configuration of the linkage group, due to the presence of the translocation.Based on these models, the relationships between the relative frequencies of the different classes of gametes are predicted for several theoretically possible arrangements of the genes within the linkage group.The models have general application for the localization of gametophytic genes and for the localization of genes in branched linkage groups.It turns out to be impossible to give an estimate for the disadvantage of ga ^- as compared to ga ⁺ from the backcross data alone. The empirical data permit only a combined estimate of the probabilities of fertilization by ga ^- and of crossing over between Ga and the locus of the marker gene.Comparison between the empirical data and the model makes it possible to determine the location of the marker loci and the locus of Ga with respect to one another. A branched linkage group appears most likely, but a linear arrangement in two chromosome arms cannot be ruled out.The observed cross-over values, and consequently, the map distances between the genes studied, are highly variable, both in different hybrids and in the progeny of a single hybrid.

---

---

Angenommen durch G. Melchers und W. Seyffert     

CLONAL VARIATION OF GRAPE-STEM AND PHYLLOXERA-GALL CALLUS GROWING IN VITRO IN DIFFERENT CONCENTRATIONS OF SUGARS

Arya , H. C., A. C. Hildebrandt , and A. J. Riker . (University of Wisconsin, Madison, Wisconsin.) Clonal variation of grape-stem and Phylloxera-gall callus growing in vitro in different concentrations of sugars. Amer. Jour. Bot. 49(4): 368–372. Illus. 1962.—The original callus grown from normal tissue and that grown from gall tissue contained mixtures of different kinds of cells. To study the variability, a large number of clones were developed by single-cell technique. From these, 6 clones were selected for detailed study. Growth was compared of 6 single-cell clones established in vitro; 3 from normal grape stem and 3 from gall tissues incited by Phylloxera vastatrix Planch. The clones were stable in growth rate (fast, medium, and slow) when grown on modified White's basal medium supplemented with coconut milk, α-naphthaleneacetic acid (NAA), and calcium pantothenate. Growth was measured after 6 weeks as the average wet weight on concentrations from 0.06–4.0% of sucrose, d(+) dextrose, d(-) levulose, d(+) mannose, d(+) galactose, and d(+) lactose, respectively. Every sugar, except mannose, was a suitable source of carbon. The cells were not all alike in their growth response to different sugars. Gall- and normal-tissue clones grew best with 0.125% sucrose, levulose, and galactose. With dextrose and lactose, optimum yields were obtained at the 1.0% sugar level. Growth of fast-, medium-, and slow-growing clones was altered with the type of sugar. Gall and normal tissues were differentiated from each other when grown on mannose in which gall tissues grew best at 0.125% and normal at 1.0% levels. Gall tissues as a group were able to grow better with mannose than the corresponding normal ones. Levulose, on the other hand, favored growth of normal clones in comparison to that of diseased ones. Sugars varied in their inhibitory influence at the 4.0% level. Dextrose, levulose, and lactose at 1.0% proved better than sucrose for the growth of all except one fast-growing gall clone which grew best with sucrose. However, at 0.125% sugar levels, even in cases where high yields were obtained, the physical character of the tissues changed to dry, brownish, and very friable. Thus, the original callus from normal and gall tissues contained cells with diverse characteristics. The various clones developed used the same sugars but varied strikingly in the rate and type of growth on certain sugars.     

Psychological interventions to reduce positive symptoms in schizophrenia: systematic review and network meta‐analysis

Psychological treatments are increasingly regarded as useful interventions for schizophrenia. However, a comprehensive evaluation of the available evidence is lacking and the benefit of psychological interventions for patients with current positive symptoms is still debated. The present study aimed to evaluate the efficacy, acceptability and tolerability of psychological treatments for positive symptoms of schizophrenia by applying a network meta‐analysis approach, that can integrate direct and indirect comparisons. We searched EMBASE, MEDLINE, PsycINFO, PubMed, BIOSIS, Cochrane Library, World Health Organization's International Clinical Trials Registry Platform and ClinicalTrials.gov for randomized controlled trials of psychological treatments for positive symptoms of schizophrenia, published up to January 10, 2018. We included studies on adults with a diagnosis of schizophrenia or a related disorder presenting positive symptoms. The primary outcome was change in positive symptoms measured with validated rating scales. We included 53 randomized controlled trials of seven psychological interventions, for a total of 4,068 participants receiving the psychological treatment as add‐on to antipsychotics. On average, patients were moderately ill at baseline. The network meta‐analysis showed that cognitive behavioural therapy (40 studies) reduced positive symptoms more than inactive control (standardized mean difference, SMD=?0.29; 95% CI: –0.55 to ?0.03), treatment as usual (SMD=?0.30; 95% CI: –0.45 to ?0.14) and supportive therapy (SMD=?0.47; 95% CI: –0.91 to ?0.03). Cognitive behavioural therapy was associated with a higher dropout rate compared with treatment as usual (risk ratio, RR=0.74; 95% CI: 0.58 to 0.95). Confidence in the estimates ranged from moderate to very low. The other treatments contributed to the network with a lower number of studies. Results were overall consistent in sensitivity analyses controlling for several factors, including the role of researchers’ allegiance and blinding of outcome assessor. Cognitive behavior therapy seems to be effective on positive symptoms in moderately ill patients with schizophrenia, with effect sizes in the lower to medium range, depending on the control condition.