A plasmid-based expression system to study protein–protein interactions at the Golgi in vivo

There is still an unmet need for simple methods to verify, visualize, and confirm protein–protein interactions in vivo. Here we describe a plasmid-based system to study such interactions. The system is based on the transmembrane domain (TMD) of the EF-hand Ca²⁺ sensor protein calneuron-2. We show that fusion of 28 amino acids that include the TMD of calneuron-2 to proteins of interest results in prominent localization on the cytoplasmic side of the Golgi. The recruitment of binding partners to the protein of interest fused to this sequence can then be easily visualized by fluorescent tags.     

Are certain life histories particularly prone to local extinction?

Using stochastic simulations and elasticity analysis, we show that there are inherent differences in the risk of extinction between life histories with different demographies. Which life history is the most vulnerable depends on which vital rate varies. When juvenile survival varies semelparous organisms with delayed reproduction are the most vulnerable ones, while a varying developmental rate puts a greater threat to semelparous organisms with rapid development. Iteroparous organisms are the most vulnerable ones when adult survival varies. Generally, we do not expect to observe organisms in nature having variation in vital rates that produce a high risk of extinction. Given the results here we therefore predict that iteroparous organisms should show low variation in adult survival. Moreover, we predict that semelparous organisms should show low variation in juvenile survival and low variation in developmental rate. The effect of temporal correlation on extinction risk is most pronounced in organisms with semelparous life histories.     

Are trials in New World monkeys on the critical path for blood-stage malaria vaccine development?

The development of malaria blood-stage vaccines is gathering momentum: there are several new funding initiatives, one multiantigen formulation is currently being tested and at least one other blood-stage vaccine is expected to begin trials in 2001. However, there is no consensus over the best way to select which form of an antigen to take into clinical testing. There is thus a danger that less-effective vaccines might be tested in the field in the order of their availability, rather than merit. Here, we argue that first proving efficacy in the New World monkey challenge model would accelerate development.     

Cordia millenii: on the risk of local extinction?

Selective logging of valuable tropical timber trees is a conservation concern because it threatens the long-term sustainability of forests. However, there is insufficient information regarding the postlogging recovery of harvested species. Here, I assessed the seed dispersal patterns, recruitment and abundance of Cordia millenii , a valuable timber tree in two Ugandan tropical rain forests that have been subjected to varying disturbance regimes. The aim was to determine the vulnerability of Cordia in these forests. The rate of seed dispersal was lower in the heavily disturbed Mabira Forest compared with the less disturbed Budongo Forest. Frugivores in Mabira were small-bodied individuals that spat seeds beneath fruiting trees, whereas 90% of the fruit in Budongo was consumed by large-bodied chimpanzees that disperse seeds over long distances. Juveniles of Cordia were not found in the closed forest, although they were found in forest gaps in Budongo but not Mabira. Mature tree density was higher in Budongo compared with Mabira. Lack of effective seed dispersal coupled with the inability of seedlings of Cordia to establish under closed canopy account for the arrested recruitment in Mabira. Enrichment planting in felling gaps is necessary to avoid local extinction of Cordia in forests without large vertebrates.     

Patient Parents: Do Offspring Decide on the Timing of Fledging in Zebra Finches?

Parent–offspring conflict over parental care is predicted to become most pronounced during offspring transition to independence when offspring are predicted to attempt to extend care for longer than parents are selected to provide it. However, on the proximate level, it is difficult to determine who plays the most important role in this process, parents or offspring. For several vertebrate taxa, it has been documented that parents end brood care by abandoning offspring after a fixed period or else show high flexibility in the duration of care, but teasing apart the role of offspring and parents underlying this flexibility has been difficult. Here, we studied the decision to fledge in captive zebra finches (Taeniopygia guttata), an altricial songbird. We experimentally delayed the time of fledging to determine who decides about the end of feeding inside the nest, parents or offspring. The experiment indicates that parents do not primarily rely on phenotypic offspring traits in their decision to feed offspring in the nest, but appear to adjust the duration of parental care as long as offspring are in the nest which parents may take as an indicator of offspring need and locomotor abilities. Delayed‐fledging offspring appeared not to suffer a disadvantage in terms of age at the onset of independent feeding. Our study suggests that, in zebra finches, offspring play a major role in determining the time of fledging and leave the nest on their own, possibly to reduce the risk of nest predation, or to evade sibling competition in the nest.     

Are we protected? Imported measles - on the way to eradication

In accordance with the 2015 regional goal for measles and rubella elimination of the WHO European Region, only a few imported cases have been documented of both diseases in Hungary for years.This paper presents a case of a Hungarian woman, born in 1975, who received measles vaccination at age of 12 months and later at age of 11 years, according to her certificate of vaccination. In 2009, after arriving home from a vacation in Ireland, she developed acute measles infection with clinical symptoms. It was confirmed by the detection of measles specific IgM, IgA and IgG antibodies, and by detection of viral nucleic acid from throat swab in virus transport medium.Additionally, an outbreak occurred in December of 2011 among a family emigrated from Romania to Hungary. No new measles cases were diagnosed among the contact persons of neither the young Hungarian woman returning from Ireland, nor the family emigrated from Romania. This observation refers to the effectiveness of the Hungarian vaccination program.     

From the first to the second domain of gelsolin: a common path on the surface of actin?

We present the 2.6 A resolution crystal structure of a complex formed between G-actin and gelsolin fragment Met25-Gln160 (G1+). The structure differs from those of other gelsolin domain 1 (G1) complexes in that an additional six amino acid residues from the crucial linker region into gelsolin domain 2 (G2) are visible and are attached securely to the surface of actin. The linker segment extends away from G1 up the face of actin in a direction that infers G2 will bind along the same long-pitch helical strand as the actin bound to G1. This is consistent with a mechanism whereby G2 attaches gelsolin to the side of a filament and then directs G1 toward a position where it would disrupt actin-actin contacts. Alignment of the sequence of the structurally important residues within the G1-G2 linker with those of WH2 (WASp homology domain 2) domain protein family members (e.g. WASp (Wiscott-Aldridge syndrome protein) and thymosin beta4) suggests that the opposing activities of filament assembly and disassembly may exploit a common patch on the surface of actin.     

How catastrophic was the end‐Triassic mass extinction?

A review of marine and terrestrial animal and plant fossils fails to reveal convincing evidence of a global catastrophe at the Triassic-Jurassic boundary, although this time marked the final disappearance of ceratite ammonites and conodonts, together with the extinction of most calcareous demosponges; important groups of bivalves and brachiopods went extinct. Because of facies problems, however, there is no stratigraphic section that reveals a clear-cut disappearance over a short distance. Other marine animal groups except perhaps the radiolarians fail to reveal a notable extinction of global extent immediately across the boundary. On the other hand, there was a substantially higher extinction rate among marine animals in the Rhaetian as compared with the previous stage. On the land, the record is equivocal. Dramatic changes across the T-J boundary have been claimed for plants in particular areas, such as eastern North America and East Greenland, but only gradual change has been recognized elsewhere. Similarly, claims of a T-J boundary vertebrate mass extinction have not been supported by others. For the Rhaetian as a whole, however, the turnover rate of reptiles was high. Although much remains to be learned, it seems evident that the fossil record of the latest Triassic is more consistent with a gradual scenario extended over time than a 'geologically instantaneous' impact catastrophe.     

Where is the right path heading from the centromere to spindle microtubules?

The kinetochore is a large protein complex that ensures accurate chromosome segregation during mitosis by connecting the centromere and spindle microtubules. One of the kinetochore sub-complexes, the constitutive centromere-associated network (CCAN), associates with the centromere and recruits another sub-complex, the KMN (KNL1, Mis12, and Ndc80 complexes) network (KMN), which binds to spindle microtubules. The CCAN-KMN interaction is mediated by two parallel pathways (CENP-C- and CENP-T-pathways) in the kinetochore, which bridge the centromere and microtubules. Here, we discuss dynamic protein-interaction changes in the two pathways that couple the centromere with spindle microtubules during mitotic progression.     

‘Accelerated aging’: a primrose path to insight?

Organism envy afflicts most researchers who work on aging in mice; how frustrating it is to see the worm and fly biologists nail down milestone after milestone, citation after citation! Surely genetic trickery can produce mice that age in a comparable jiffy? Alas, our near‐total ignorance of what times the aging process makes it hard to guess what genes to tweak, if indeed aging can be mimicked a presto. Building a case that a given short‐lived mutant ages quickly is a steep and thorny path, requiring more than just plucking a symptom here and there from a list of things that sometimes go wrong in old people or old mice. The hallmark of aging is that a lot goes wrong more or less at the same time, in 2‐year‐old mice, 10‐year‐old dogs and 70‐year‐old people. Finding ways to damage one or two systems in a 6‐week or 6‐month‐old mouse is not too hard to do, but the implications of such studies for improved understanding of aging per se are at best indirect and at worst imaginary and distracting.     

Altering the 3′ UTR endonucleolytic cleavage site of a Chlamydomonas chloroplast mRNA affects 3′-end maturation in vitro but not in vivo

The 3 ends of chloroplast mRNAs are produced by the processing of longer precursors. The 3 ends of most plastid mRNAs are located at, or several nucleotides downstream of, stem-loop structures, which act as 3-end-processing signals and RNA stability elements. In chloroplasts of the green alga Chlamydomonas reinhardtii, 3-end maturation of atpB mRNA involves endonucleolytic cleavage of the pre-mRNA at an AU-rich site located about 10 nucleotides downstream of the stem-loop structure. This cleavage is followed by exonucleolytic resection to generate the mature 3 end. In order to define critical nucleotides of the endonucleolytic cleavage site, we mutated its sequence. Incubation of synthetic atpB pre-RNAs containing these mutations in a chloroplast protein extract resulted in the accumulation of 3-end-processed products. However, in two cases where the AU-rich sequence of this site was replaced with a GC-rich one, the 3 end of the stable processing product differed from that of the wild-type product. To examine whether these mutations affected atpB mRNA processing or accumulation in vivo, the endogenous 3 UTR was replaced with mutated sequences by biolistic transformation of Chlamydomonas chloroplasts. Analysis of the resulting strains revealed that the accumulation of atpB mRNA was approximately equal to that of wild-type cells, and that a wild-type atpB 3 end was generated. These results imply that Chlamydomonas atpB 3 processing parallels the situation with other endonucleases such as Escherichia coli RNAse E, where specific sequences are required for correct in vitro processing, but in vivo these mutations can be overcome.     

Species’ response patterns to habitat fragmentation: do trees support the extinction threshold hypothesis?

In fragmented landscapes the relationship between the probability of occurrence of single species and the amount of suitable habitat is usually not proportional, with a threshold habitat level below which the population becomes extinct. Ecological theory predicts that, although the reduction in species’ occurrence probabilities (and eventually the extinction threshold) is a direct consequence of habitat loss, habitat fragmentation might reduce species’ occurrence probabilities and affect the location of this threshold by reducing its predicted occurrence to lower levels of habitat amount. However, little is known about the validity of this extinction threshold hypothesis. Here, we performed analyses on the relationships between the probability of occurrence of eight tree species and the availability of forest habitat for two different empirical scenarios of low and moderate to high fragmentation. We partitioned the effects of habitat amount versus fragmentation by using two metrics: (1) the percentage of forest cover, and (2) the proportion of this percentage occurring in the largest forest patch. We find that, although decreasing forest cover had negative effects on the occurrence of tree species irrespective of fragmentation levels, forest fragmentation significantly modified the response pattern in six tree species, although only one species confirmed the extinction threshold hypothesis, which we interpret as a consequence of high degree of forest specialism and low dispersal ability. For most species, fragmentation either had positive effects or did not affect significantly the species’ probability of occurrence. This indicates that the effects of habitat fragmentation on tree species are weak relative to the effects of habitat amount, which is the main determinant of the reduction in species’ occurrence probabilities, and eventually species extinction, in fragmented landscapes.     

Plant natural products: Back to the future or into extinction?

Natural product substances have historically served as the most significant source of new leads for pharmaceutical development. However, with the advent of robotics, bioinformatics, high throughput screening (HTS), molecular biology-biotechnology, combinatorial chemistry, in silico (molecular modeling) and other methodologies, the pharmaceutical industry has largely moved away from plant derived natural products as a source for leads and prospective drug candidates. Can, or will, natural products ever recapture the preeminent position they once held as a foundation for drug discovery and development? The challenges associated with development of natural products as pharmaceuticals are illustrated by the Taxol^® story. Several misconceptions, which constrain utilization of plant natural products, for discovery and development of pharmaceuticals, are addressed to return natural products to the forefront.     

Are comparative studies of extinction risk useful for conservation?

Large-scale, comparative studies of species extinction risk have become common in conservation science, but their influence on conservation practice appears limited. The link between such studies and the practice of conservation breaks down in two key places. First, results of comparative studies are often ambiguous, inconsistent and difficult to translate into policy. Second, conservation as currently practiced emphasizes the rescue and protection of currently threatened biodiversity, whereas comparative studies are often better suited to a proactive approach that anticipates and prevents future species declines. Scientists should make their research more accessible by addressing the first issue. Policymakers and managers, in turn, could make better use of comparative studies by moving towards more preventative approaches to conservation planning.     

Are α-adrenergic antagonists potent tocolytics? In vivo experiments on postpartum rats

The aim of this study was to investigate the effects of α-adrenergic antagonists on the motor activity of the postpartum uterus of the rat in vivo. Intrauterine pressure was assessed by means of a Millar catheter fitted with a latex microballoon. Some of the tested compounds (urapidil, yohimbine, phentolamine, benoxathian and prazosin) decreased the uterine activity to a significant extent (57.4–67.4%). However, none of the investigated α-receptor blockers exerted the same effect as β-adrenergic agonists. Our results suggest that α-adrenergic antagonists could possibly be used as an alternative to β-adrenergic agonists in clinical tocolysis after an appropriate clinical evaluation.