Identification and characterization of a novel Bacillus thuringiensis δ-endotoxin entomocidal to coleopteran and lepidopteran larvae

A new class of Bacillus thuringiensis delta-endotoxins, or insecticidal control proteins (ICPs), is defined by an apparently cryptic protein with a unique primary structure and novel entomocidal specificity for certain coleopteran and lepidopteran species. The discovery of a new group of ICPs will extend the use of this natural insecticide in integrated pest-management systems.     

Glyoxal as an alternative fixative to formaldehyde in immunostaining and super‐resolution microscopy

Paraformaldehyde (PFA) is the most commonly used fixative for immunostaining of cells, but has been associated with various problems, ranging from loss of antigenicity to changes in morphology during fixation. We show here that the small dialdehyde glyoxal can successfully replace PFA. Despite being less toxic than PFA, and, as most aldehydes, likely usable as a fixative, glyoxal has not yet been systematically tried in modern fluorescence microscopy. Here, we tested and optimized glyoxal fixation and surprisingly found it to be more efficient than PFA‐based protocols. Glyoxal acted faster than PFA, cross‐linked proteins more effectively, and improved the preservation of cellular morphology. We validated glyoxal fixation in multiple laboratories against different PFA‐based protocols and confirmed that it enabled better immunostainings for a majority of the targets. Our data therefore support that glyoxal can be a valuable alternative to PFA for immunostaining.     

Testing top‐down and bottom‐up effects on arid zone beetle assemblages following mammal reintroduction

Species extinctions and declines are occurring globally and commonly have cascading effects on ecosystems. In Australia, mammal extinctions have been extensive, particularly in arid areas, where precipitation drives ecosystems. Many ecologically extinct mammals feed on soil‐dwelling insects. However, how this top‐down pressure affected their prey and how this contrasts with the bottom‐up impacts of fluctuating precipitation remains unclear. We constructed a long‐term exclusion experiment in a multi‐species mammal reintroduction zone in semi‐arid Australia to test how top‐down (reintroduced mammals) and bottom‐up (precipitation) factors affect root‐feeding chafer beetles (Coleoptera: Melolonthinae). We used emergence traps in ten replicate 20 × 20 m plots of control, exclusion and procedural control treatments to trap chafers biannually from 2009 to 2015. Annual precipitation during this period varied from 173 to 481 mm. Mammal exclusion did not affect chafers, indicating that top‐down regulation was not important. Instead, chafer abundance, species density and biomass increased with precipitation. Chafer body size and assemblage composition were best predicted by sampling year, suggesting that random drift determined species abundances. Increased resource availability therefore favoured all species similarly. We thus found no evidence that mammal predation alters chafer populations and conclude that they may be driven primarily by bottom‐up processes. Further research should determine if the cascading effects of species loss are less important for herbivores generally than for higher level trophic groups and the role of ecosystem stability in mediating these patterns.     

Phorbol ester and diacylglycerol activation of native protein kinase C species from various tissues

The characteristics of PKC activation induced by a number of compounds were investigated using PKCs, partially-purified from sources with a naturally high abundance of certain Ca²⁺ dependent PKC isoforms. Native isoforms were used rather than PKC isoforms expressed from a baculovirus system to assess the effect of tissue specific factors on activity. However, some data using recombinant PKC were included for comparison.The presence of specific PKC isoforms in different tissues was determined using Western blot analysis. Protein kinase C , ₁, , , and / were all present in rat midbrain cytosolic extract, PKC , ₁, , and / were present in spleen cytosol, and PKC and / were present in COS 7 cell cytosol. The predominance of and activities in COS 7 and spleen extracts respectively was confirmed by enzymic assay.The PKC activity assay was configured such that the Ca²⁺ dependence of the PKC activity induced by different PKC activators could be determined. Phorbol 12,13-dibutyrate (PDBu) was virtually equipotent on the Ca²⁺-dependent PKC activity from midbrain and spleen and slightly less potent on that from COS 7 cells. In the absence of Ca²⁺, PDBu was considerably less potent overall (as, indeed, were the other PKC activators) and was less potent on COS 7 cell PKC than on those from midbrain or spleen. Mezerein was more potent than PDBu at inducing PKC activity in COS 7 cell extracts in either the absence or presence of Ca²⁺ whereas in the presence of Ca²⁺, mezerein was slightly less potent on midbrain and spleen than PDBu and equipotent in the absence of Ca²⁺. Maximum values for Ca²⁺-independent activation by mezerein indicated that this activator was particularly effective in recruiting Ca²⁺-dependent PKC isoform activity in a Ca²⁺ free environment. The greater potency of mezerein on PKC was confirmed using PKC and further purified from rat spleen by hydroxylapatite (HAP) chromatography. The effects of both PDBu and mezerein were investigated using anterior pituitary tissue where a particularly high potency of mezerein in the absence of Ca²⁺ was noted. The diacylglycerol, 1,2-dioctanoyl-sn-glycerol (DOG), appeared to cause little or no activation of native Ca²⁺-dependent isoforms in Ca²⁺ free conditions unlike another longer chain diacylglycerol, 1,2-dioleoyl-sn-glycerol. Also DOG activated midbrain PKCs more potently than PKCs from spleen or COS 7 cells (or lung and pituitary tissue) in the presence of Ca²⁺. The concentration-dependence of DOG was examined on PKC and PKC further purified from brain by HAP chromatography, revealing that DOG was equally potent on both of these isoforms derived from brain and on recombinant PKC . However, [³H]PDBu binding data using PKC purified from several sources gave very different IC₅₀ values when DOG was used as a displacer, and in general these values correlated with the EC₅₀ values recorded from the activity assay.The data presented here indicate that there are distinct differences in the activator pharmacology of different native PKC isoforms and between the same isoform expressed in different tissues, either because of post-translational modifications or some other tissue specific factor.     

Turning the tables of sex distinction in craniofacial identification: Why females possess thicker facial soft tissues than males,not vice versa

Males are universally reported to possess larger facial soft‐tissue thickness (FSTT) than females, however, this observation oversimplifies the raw data yielding an underpowered assessment of FSTT sex‐patterning where: differences are small (η² < 5%) and inconsistent (females are routinely larger than males at the cheeks). Here we investigate body‐size normalized data to assess whether more general and improved understanding of FSTT sex‐variation in humans is possible. FSTTs were measured in 52 healthy living Australians aged 18 to 30 years using B‐mode ultrasound. Participants' stature and body mass were also measured. Sex differences were calculated before and after normalization by the aforementioned body‐composition variables. Methods were repeated in three other independent samples to evaluate reproducibility: 100 American Whites and 60 American Blacks measured by B‐mode ultrasound; and 50 Turkish residents measured by regular supine CT. Compared to raw mean differences (F < M, by ?6%), females displayed much thicker FSTTs than males when normalized for body mass (F > M, by +16%). Consequently, while the sexes share similar raw values, females possess much larger FSTTs for their relatively lighter bodies. The relative FSTT difference was 2.7× larger than the raw mean difference. Sex differences in FSTT are of larger magnitude and reversed direction in mass normalized data. Contrary to popular thought, females possess much larger FSTTs than males owing to their generically lighter bodies (?18 kg). These data patterns help explain why the pooling of sex‐categorized FSTT does not jeopardize the sex‐difference—it is encoded more strongly in terms relative to body mass.     

Risks and Population Burden of Cardiovascular Diseases Associated with Diabetes in China: A Prospective Study of 0.5 Million Adults

BackgroundIn China, diabetes prevalence is rising rapidly, but little is known about the associated risks and population burden of cardiovascular diseases. We assess associations of diabetes with major cardiovascular diseases and the relevance of diabetes duration and other modifiable risk factors to these associations.ConclusionsAmong Chinese adults, diabetes is associated with significantly increased risks of major cardiovascular diseases. The increasing prevalence and younger age of onset of diabetes foreshadow greater diabetes-attributable disease burden in China.     

World Health Organization Estimates of the Relative Contributions of Food to the Burden of Disease Due to Selected Foodborne Hazards: A Structured Expert Elicitation

Background

The Foodborne Disease Burden Epidemiology Reference Group (FERG) was established in 2007 by the World Health Organization (WHO) to estimate the global burden of foodborne diseases (FBDs). This estimation is complicated because most of the hazards causing FBD are not transmitted solely by food; most have several potential exposure routes consisting of transmission from animals, by humans, and via environmental routes including water. This paper describes an expert elicitation study conducted by the FERG Source Attribution Task Force to estimate the relative contribution of food to the global burden of diseases commonly transmitted through the consumption of food.

Methods and Findings

We applied structured expert judgment using Cooke’s Classical Model to obtain estimates for 14 subregions for the relative contributions of different transmission pathways for eleven diarrheal diseases, seven other infectious diseases and one chemical (lead). Experts were identified through international networks followed by social network sampling. Final selection of experts was based on their experience including international working experience. Enrolled experts were scored on their ability to judge uncertainty accurately and informatively using a series of subject-matter specific ‘seed’ questions whose answers are unknown to the experts at the time they are interviewed. Trained facilitators elicited the 5^th, and 50^th and 95^th percentile responses to seed questions through telephone interviews. Cooke’s Classical Model uses responses to the seed questions to weigh and aggregate expert responses. After this interview, the experts were asked to provide 5^th, 50^th, and 95^th percentile estimates for the ‘target’ questions regarding disease transmission routes. A total of 72 experts were enrolled in the study. Ten panels were global, meaning that the experts should provide estimates for all 14 subregions, whereas the nine panels were subregional, with experts providing estimates for one or more subregions, depending on their experience in the region. The size of the 19 hazard-specific panels ranged from 6 to 15 persons with several experts serving on more than one panel. Pathogens with animal reservoirs (e.g. non-typhoidal Salmonella spp. and Toxoplasma gondii) were in general assessed by the experts to have a higher proportion of illnesses attributable to food than pathogens with mainly a human reservoir, where human-to-human transmission (e.g. Shigella spp. and Norovirus) or waterborne transmission (e.g. Salmonella Typhi and Vibrio cholerae) were judged to dominate. For many pathogens, the foodborne route was assessed relatively more important in developed subregions than in developing subregions. The main exposure routes for lead varied across subregions, with the foodborne route being assessed most important only in two subregions of the European region.

Conclusions

For the first time, we present worldwide estimates of the proportion of specific diseases attributable to food and other major transmission routes. These findings are essential for global burden of FBD estimates. While gaps exist, we believe the estimates presented here are the best current source of guidance to support decision makers when allocating resources for control and intervention, and for future research initiatives.