Abundance,density and activity of Salvator merianae (Reptilia: Teiidae) and the effect of poaching on the site occupancy by the lizard in an Atlantic Forest Reserve,Brazil

We aimed to estimate the density, occupancy and detectability of Salvator merianae (Tegu) in one of the largest Atlantic rainforest remnants in Espírito Santo, Brazil, the VNR. Species patch occupancy was modelled and used to predict the response direction of six covariates based on prior knowledge of the Tegu's ecology. A priori, we expected that the covariates measured should represent key habitat features for the species (i.e. temperature, forest edge, open habitats) or elements possibly avoided by the species, based on the hypothesis that poaching would have a negative effect on patch occupancy. We used line‐transect surveys to estimate density and abundance. Camera‐traps were used to estimate patch occupancy by the Tegu. Estimated density for S. merianae was 0.21 ± 0.02 Tegus/ha and estimated population size was 4990 ± 521 individuals. Patch occupancy was best described by two covariates: poaching intensity and distance to the forest edge. Detectability was affected by three covariates: poaching intensity, tree density and temperature. Our study presents robust information on abundance and density, habitat use, and activity of S. merianae in the VNR and is the first study providing data on the effects that poaching has on patch occupancy of this lizard. The data indicated that the occupancy and detectability of this species were influenced by a set of factors, providing information that can be useful in management plans in areas where this species can potentially decline and in areas where it may be introduced.     

Neurobeachin regulates neurotransmitter receptor trafficking to synapses

The surface density of neurotransmitter receptors at synapses is a key determinant of synaptic efficacy. Synaptic receptor accumulation is regulated by the transport, postsynaptic anchoring, and turnover of receptors, involving multiple trafficking, sorting, motor, and scaffold proteins. We found that neurons lacking the BEACH (beige-Chediak/Higashi) domain protein Neurobeachin (Nbea) had strongly reduced synaptic responses caused by a reduction in surface levels of glutamate and GABA_A receptors. In the absence of Nbea, immature AMPA receptors accumulated early in the biosynthetic pathway, and mature N-methyl-d-aspartate, kainate, and GABA_A receptors did not reach the synapse, whereas maturation and surface expression of other membrane proteins, synapse formation, and presynaptic function were unaffected. These data show that Nbea regulates synaptic transmission under basal conditions by targeting neurotransmitter receptors to synapses.     

Adipose triglyceride lipase is a TG hydrolase of the small intestine and regulates intestinal PPARα signaling

Adipose triglyceride lipase (ATGL) is the rate-limiting enzyme mediating triglyceride (TG) hydrolysis. The lack of ATGL results in TG accumulation in multiple tissues, underscoring the critical role of ATGL in maintaining lipid homeostasis. Recent evidence suggests that ATGL affects TG metabolism via activation of peroxisome proliferator-activated receptor α (PPARα). To investigate specific effects of intestinal ATGL on lipid metabolism we generated mice lacking ATGL exclusively in the intestine (ATGLiKO). We found decreased TG hydrolase activity and increased intracellular TG content in ATGLiKO small intestines. Intragastric administration of [³H]trioleate resulted in the accumulation of radioactive TG in the intestine, whereas absorption into the systemic circulation was unchanged. Intraperitoneally injected [³H]oleate also accumulated within TG in ATGLiKO intestines, indicating that ATGL mobilizes fatty acids from the systemic circulation absorbed by the basolateral side from the blood. Down-regulation of PPARα target genes suggested modulation of cholesterol absorption by intestinal ATGL. Accordingly, ATGL deficiency in the intestine resulted in delayed cholesterol absorption. Importantly, this study provides evidence that ATGL has no impact on intestinal TG absorption but hydrolyzes TGs taken up from the intestinal lumen and systemic circulation. Our data support the role of ATGL in modulating PPARα-dependent processes also in the small intestine.     

Characterization of Hepatitis C Virus Intra- and Intergenotypic Chimeras Reveals a Role of the Glycoproteins in Virus Envelopment

Hepatitis C virus (HCV) is highly variable and associated with chronic liver disease. Viral isolates are grouped into seven genotypes (GTs). Accumulating evidence indicates that viral determinants in the core to NS2 proteins modulate the efficiency of virus production. However, the role of the glycoproteins E1 and E2 in this process is currently poorly defined. Therefore, we constructed chimeric viral genomes to explore the role of E1 and E2 in HCV assembly. Comparison of the kinetics and efficiency of particle production by intragenotypic chimeras highlighted core and p7 as crucial determinants for efficient virion release. Glycoprotein sequences, however, had only a minimal impact on this process. In contrast, in the context of intergenotypic HCV chimeras, HCV assembly was profoundly influenced by glycoprotein genes. On the one hand, insertion of GT1a-derived (H77) E1-E2 sequences into a chimeric GT2a virus (Jc1) strongly suppressed virus production. On the other hand, replacement of H77 glycoproteins within the GT1a-GT2a chimeric genome H77/C3 by GT2a-derived (Jc1) E1-E2 increased infectious particle production. Thus, within intergenotypic chimeras, glycoprotein features strongly modulate virus production. Replacement of Jc1 glycoprotein genes by H77-derived E1-E2 did not grossly affect subcellular localization of core, E2, and NS2. However, it caused an accumulation of nonenveloped core protein and increased abundance of nonenveloped core protein structures with slow sedimentation. These findings reveal an important role for the HCV glycoproteins E1 and E2 in membrane envelopment, which likely depends on a genotype-specific interplay with additional viral factors.     

Reprogramming the Phenylpropanoid Metabolism in Seeds of Oilseed Rape by Suppressing the Orthologs of REDUCED EPIDERMAL FLUORESCENCE1

As a result of the phenylpropanoid pathway, many Brassicaceae produce considerable amounts of soluble hydroxycinnamate conjugates, mainly sinapate esters. From oilseed rape (Brassica napus), we cloned two orthologs of the Arabidopsis (Arabidopsis thaliana) gene REDUCED EPIDERMAL FLUORESCENCE1 (REF1) encoding a coniferaldehyde/sinapaldehyde dehydrogenase. The enzyme is involved in the formation of ferulate and sinapate from the corresponding aldehydes, thereby linking lignin and hydroxycinnamate biosynthesis as a potential branch-point enzyme. We used RNA interference to silence REF1 genes in seeds of oilseed rape. Nontargeted metabolite profiling showed that BnREF1-suppressing seeds produced a novel chemotype characterized by reduced levels of sinapate esters, the appearance of conjugated monolignols, dilignols, and trilignols, altered accumulation patterns of kaempferol glycosides, and changes in minor conjugates of caffeate, ferulate, and 5-hydroxyferulate. BnREF1 suppression affected the level of minor sinapate conjugates more severely than that of the major component sinapine. Mapping of the changed metabolites onto the phenylpropanoid metabolic network revealed partial redirection of metabolic sequences as a major impact of BnREF1 suppression.Phenylpropanoid metabolism provides plants with a vast array of phenolic compounds that contribute to nearly all aspects of plant life (Vogt, 2010). In species of the Brassicaceae family, soluble hydroxycinnamate conjugates, mainly sinapate esters, constitute an abundant metabolite fraction produced from a branch of the phenylpropanoid pathway (Fraser and Chapple, 2011). As major compounds, sinapoylmalate accumulates in leaves (Hause et al., 2002) and sinapoylcholine (sinapine) in seeds (Bouchereau et al., 1991; Fig. 1).Open in a separate windowFigure 1.Biosynthesis of sinapate esters emphasizing the crucial role of the bifunctional hydroxycinnamaldehyde dehydrogenase CALDH/SALDH encoded by the gene REF1. Dashed arrows symbolize multistep biosyntheses. Abbreviations for enzymes are as follows: CAD, (hydroxy)cinnamyl alcohol dehydrogenase; SAD, sinapyl alcohol dehydrogenase; SCT, 1-O-sinapoylglucose:choline sinapoyltransferase; SGT, UDP-Glc:sinapate glucosyltransferase; SMT, 1-O-sinapoylglucose:malate sinapoyltransferase.In Arabidopsis (Arabidopsis thaliana), the disturbed accumulation of sinapoylmalate caused the mutant phenotype reduced epidermal fluorescence (ref; Ruegger and Chapple, 2001). Molecular characterization of the ref1 mutant led to the identification of the gene At3g24503 (REF1) encoding coniferaldehyde dehydrogenase/sinapaldehyde dehydrogenase (CALDH/SALDH; EC 1.2.1.68; Nair et al., 2004). The bifunctional enzyme CALDH/SALDH was shown to catalyze the NADP⁺-dependent oxidation of coniferaldehyde and sinapaldehyde to yield the corresponding hydroxycinnamates ferulate and sinapate. As a potential branching enzyme, the enzymatic activity of the REF1-encoded CALDH/SALDH might be crucial for the partition ratio of metabolites between lignin and hydroxycinnamate biosynthesis (Fig. 1). Therefore, manipulation of REF1 expression coupled to a comprehensive metabolite analysis of transgenic plants appeared as an interesting strategy to gain a deeper understanding of the plant phenylpropanoid metabolic network. Moreover, in crop plants like oilseed rape (Brassica napus), where high levels of sinapate esters contribute to antinutritive features, the suppression of REF1 orthologs might cause increased quality.This work describes the isolation of REF1 orthologous genes from oilseed rape (BnREF1). By RNA interference, transgenic lines of oilseed rape were generated that suppress BnREF1 gene expression during seed development. Homozygous transgenic progeny were developed and used in a comprehensive liquid chromatography-mass spectrometry (LC/MS)-based metabolite profiling to investigate the impact of BnREF1 silencing in seeds. Mapping of changed metabolite patterns onto the phenylpropanoid metabolic network revealed insight into the molecular mechanisms by which silencing of BnREF1 produced a novel seed chemotype in oilseed rape.     

The impact of plant diversity and fertilization on fitness of a generalist grasshopper

In many environments land use intensification is likely to result in a decrease in species richness and in an increase in eutrophication. Although the importance of both factors for higher trophic levels such as insect herbivores is well documented, their impact has rarely been studied in combination. Herbivorous insects have a strong impact on the functioning of ecosystems and it is therefore important to understand how they are affected by eutrophication in high or low diversity environments.We used a grassland biodiversity experiment to investigate the combined effect of fertilization and plant diversity loss on the fitness of the generalist grasshopper Chorthippus parallelus by rearing grasshopper nymphs for four weeks in cages on unfertilized or fertilized (NPK) subplots across a species richness gradient from 1 to 60 plant species. Survival, the number of oothecae, body mass and the number of hatchlings were measured separately for each cage. Plant diversity had no effect on any of the grasshopper fitness measures, neither in unfertilized nor in fertilized plots. NPK-fertilization reduced grasshopper survival but increased body mass of males and reproductive success of the surviving females. Fertilization effects were not mediated by plant community structure, productivity or composition, suggesting that higher food plant quality was one of the main drivers. There was no interaction between plant diversity and fertilization on any of the measures. In conclusion, an increase in eutrophication, in both species-rich and species-poor grasslands, could lead to higher reproductive success and therefore higher abundances of herbivorous insects including insect pests, with fertilization effects dominating plant diversity effects.     

A Circle-Monitor for Computerised Assessment of Visual Neglect in Peripersonal Space

Current assessment of visual neglect involves paper-and-pencil tests or computer-based tasks. Both have been criticised because of their lack of ecological validity as target stimuli can only be presented in a restricted visual range. This study examined the user-friendliness and diagnostic strength of a new “Circle-Monitor” (CM), which enlarges the range of the peripersonal space, in comparison to a standard paper-and-pencil test (Neglect-Test, NET).

Methods

Ten stroke patients with neglect and ten age-matched healthy controls were examined by the NET and the CM test comprising of four subtests (Star Cancellation, Line Bisection, Dice Task, and Puzzle Test).

Results

The acceptance of the CM in elderly controls and neglect patients was high. Participants rated the examination by CM as clear, safe and more enjoyable than NET. Healthy controls performed at ceiling on all subtests, without any systematic differences between the visual fields. Both NET and CM revealed significant differences between controls and patients in Line Bisection, Star Cancellation and visuo-constructive tasks (NET: Figure Copying, CM: Puzzle Test). Discriminant analyses revealed cross-validated assignment of patients and controls to groups was more precise when based on the CM (hit rate 90%) as compared to the NET (hit rate 70%).

Conclusion

The CM proved to be a sensitive novel tool to diagnose visual neglect symptoms quickly and accurately with superior diagnostic validity compared to a standard neglect test while being well accepted by patients. Due to its upgradable functions the system may also be a valuable tool not only to test for non-visual neglect symptoms, but also to provide treatment and assess its outcome.     

Niemann-Pick C Disease Gene Mutations and Age-Related Neurodegenerative Disorders

Niemann-Pick type C (NPC) disease is a rare autosomal-recessively inherited lysosomal storage disorder caused by mutations in NPC1 (95%) or NPC2. Given the highly variable phenotype, diagnosis is challenging and particularly late-onset forms with predominantly neuropsychiatric presentations are likely underdiagnosed. Pathophysiologically, genetic alterations compromising the endosomal/lysosomal system are linked with age-related neurodegenerative disorders. We sought to examine a possible association of rare sequence variants in NPC1 and NPC2 with Parkinson''s disease (PD), frontotemporal lobar degeneration (FTLD) and progressive supranuclear palsy (PSP), and to genetically determine the proportion of potentially misdiagnosed NPC patients in these neurodegenerative conditions. By means of high-resolution melting, we screened the coding regions of NPC1 and NPC2 for rare genetic variation in a homogenous German sample of patients clinically diagnosed with PD (n = 563), FTLD (n = 133) and PSP (n = 94), and 846 population-based controls. The frequencies of rare sequence variants in NPC1/2 did not differ significantly between patients and controls. Disease-associated NPC1/2 mutations were found in six PD patients (1.1%) and seven control subjects (0.8%), but not in FTLD or PSP. All rare variation was detected in the heterozygous state and no compound heterozygotes were observed. Our data do not support the hypothesis that rare NPC1/2 variants confer susceptibility for PD, FTLD, or PSP in the German population. Misdiagnosed NPC patients were not present in our samples. However, further assessment of NPC disease genes in age-related neurodegeneration is warranted.