Isotopic niche mirrors trophic niche in a vertebrate island invader

Caution for the indiscriminate conversion of the isotopic niche into ecologic niche was recently advised. We tested the utility of the isotopic niche to answer ecological questions on oceanic islands. We compared the isotopic niches of black rats (Rattus rattus) on two islands in the Gulf of California, Mexico: Farrallón de San Ignacio (FSI) and San Pedro Mártir (SPM). Both islands maintained several species of marine birds, but FSI is devoid of terrestrial vegetation and SPM has several species of terrestrial plants. We tested the hypothesis that rats on FSI have a narrower trophic niche due to its lower diversity of food items. We predicted a smaller variance in δ¹³C and δ¹⁵N values of rat muscle on FSI, and a lower use of marine birds as food on SPM. We also examined stomach contents of rats on both islands to validate the isotopic information. Variances in δ¹³C and δ¹⁵N values of black rats were lower on FSI, and the contribution of marine birds to the diet of rats was smaller on SPM. Stomachs in most rats collected on FSI contained only one or two types of food items, mostly marine birds and terrestrial invertebrates. In contrast, stomachs with only one type of food item were rare on SPM, and in most cases they contained three or more food types. Our findings showed that isotopic variance is a good approximation for trophic niche when comparing populations with access to an assemblage of preys with contrasting biological and isotopic diversity.     

Immunocytochemical localization of DNA in synaptonemal complexes of rat and mouse spermatocytes,and of chick oocytes

The distribution of DNA in synaptonemal complexes of rat and mouse spermatocytes, and of chick oocytes was investigated by immunogold electron microscopy. Except for a few specific sites, DNA was not immunolocalized in the space between lateral elements of the complex. Some labeled fibrils connecting the lateral elements with the central element were observed associated with recombination nodules or near them. However, other labeled fibrils in the space between lateral elements did not appear to present any relationship to recombination nodules. The immunocytochemical approaches used here confirmed the presence of significant amounts of DNA in the lateral elements as previously indicated by preferential DNA staining methods. Furthermore, our findings support the view that recombination nodules are the site of chiasma formation.     

Small‐scale Farming and Grazing Reduce Regeneration of Polylepis tomentella (Rosaceae) in the Semiarid Andes of Bolivia

In the Andean highlands, Polylepis woodlands are a rare and unique ecosystem of the treeline. Although human activities have caused the loss of extensive forested areas and decreased woodland regeneration, few systematic and quantitative assessments have been carried out in Polylepis forests. This study compares differences in habitat characteristics, population structure, and reproductive output in populations of Polylepis tomentella subject to different levels of human disturbance in the south‐central Andes of Bolivia. We selected P. tomentella because the species still covers large extensions in the form of fragmented forest patches. Results show that human activities affected all the studied populations. Moderately disturbed populations exhibited a lower percentage of farmed area (0.6%) and browsed plants (4%) than strongly disturbed populations (5% and 12%, respectively). All populations exhibited similar proportion of plants with logging scars. Potassium content and canopy closure were 1.5 and 2.5 higher, respectively, in strongly disturbed populations. The density of saplings and seedlings were 75 percent and 80 percent lower in strongly disturbed populations than in moderately disturbed population, even though reproductive individuals produced twice more flowers and fruits. Our results suggest that fruit production does not limit regeneration of P. tomentella and post‐dispersal mechanisms may decrease seed germination and increase seedling mortality. Overall, strongly disturbed populations will be less likely to regenerate, leading to population decline. Conservation programs should facilitate forest recovery by promoting seedling establishment, reducing overharvesting and over‐browsing, and protecting remaining adult plants.     

Preliminary Investigation of a 64-element Capacitive Micromachined Ultrasound Transducer (CMUT) Annular Array Designed for High Intensity Focused Ultrasound (HIFU)

Background

Treatment of prostate cancer using endocavitary High Intensity Focused Ultrasound (HIFU) has become more commonplace since the first treatments in the 1990s. The gold standard HIFU strategy to treat prostate cancer is the complete thermal ablation of the entire prostate gland under real-time ultrasound (US) image guidance. A more desirable treatment and the current trend, however, is towards a focal treatment but more accurate and finely tunable thermal lesions are needed along with improved US imaging guidance. In this study, Capacitive Micromachined Ultrasound Transducer (CMUT) technology is being investigated, as they have shown recent promise for US imaging and potential to be used for HIFU therapy. They offer potential advantages over current piezoelectric designs in the context of ultrasound-guided HIFU (USgHIFU) focal therapies.

Use of a Novel Chagas Urine Nanoparticle Test (Chunap) for Diagnosis of Congenital Chagas Disease

Background

Detection of congenital T. cruzi transmission is considered one of the pillars of control programs of Chagas disease. Congenital transmission accounts for 25% of new infections with an estimated 15,000 infected infants per year. Current programs to detect congenital Chagas disease in Latin America utilize microscopy early in life and serology after 6 months. These programs suffer from low sensitivity by microscopy and high loss to follow-up later in infancy. We developed a Chagas urine nanoparticle test (Chunap) to concentrate, preserve and detect T. cruzi antigens in urine for early, non-invasive diagnosis of congenital Chagas disease.

Methodology/Principal Findings

This is a proof-of-concept study of Chunap for the early diagnosis of congenital Chagas disease. Poly N-isopropylacrylamide nano-particles functionalized with trypan blue were synthesized by precipitation polymerization and characterized with photon correlation spectroscopy. We evaluated the ability of the nanoparticles to capture, concentrate and preserve T. cruzi antigens. Urine samples from congenitally infected and uninfected infants were then concentrated using these nanoparticles. The antigens were eluted and detected by Western Blot using a monoclonal antibody against T. cruzi lipophosphoglycan. The nanoparticles concentrate T. cruzi antigens by 100 fold (western blot detection limit decreased from 50 ng/ml to 0.5 ng/ml). The sensitivity of Chunap in a single specimen at one month of age was 91.3% (21/23, 95% CI: 71.92%–98.68%), comparable to PCR in two specimens at 0 and 1 month (91.3%) and significantly higher than microscopy in two specimens (34.8%, 95% CI: 16.42%–57.26%). Chunap specificity was 96.5% (71/74 endemic, 12/12 non-endemic specimens). Particle-sequestered T. cruzi antigens were protected from trypsin digestion.

Conclusion/Significance

Chunap has the potential to be developed into a simple and sensitive test for the early diagnosis of congenital Chagas disease.     

Spontaneous binding ofYersinia enterocolitica to murine leukocytes

Twelve strains ofYersinia enterocolitica were examined for their ability to bind spontaneously to murine leukocytes. Each of eight HeLa cell invasive strains exhibited nonselective binding to peritoneal leukocytes, lymph node leukocytes, and thymocytes, whereas four noninvasive strains lacked binding properties. Like the HeLa cell invasion, the binding ofY. enterocolitica to leukocytes was much less efficient for bacteria grown at 37°C than for bacteria grown at 22°C. The binding properties were not influenced by the virulence plasmid that codes for Vwa⁺ phenotype. This leukocyte binding test is proposed as a simple assay for invasive properties ofY. enterocolitica.     

The effect of root exudates on root architecture in <Emphasis Type=Italic>Arabidopsis thaliana</Emphasis>

Root architecture is of key importance for plant nutrition and performance. It is known that root architecture is determined by genetics and environmental conditions. The aim of the present study was to evaluate if root exudation within a given plant has a role in the development of root architecture. We conducted a series of experiments using Arabidopsis thaliana Ler and Col grown with and without activated charcoal (AC). The addition of AC lowered the concentration of secondary metabolites in the growth media by more than 90%. Our results consistently showed that the addition of AC significantly decreased the number of lateral roots (38% in Ler and 27% in Col), but this decrease was compensated by an increase in the root length per unit of lateral root (83% in Ler and 96% in Col). This compensation resulted in a non-significant effect of AC on the total length of lateral roots. The effects of AC on root architecture were partially or totally reverted by the differential supplementation of root exudates from other plants of the same ecotype. Our results indicate a direct role of secondary metabolites present in the root exudates in the development of root architecture.     

The reliability of the ankle-brachial index in the Atherosclerosis Risk in Communities (ARIC) study and the NHLBI Family Heart Study (FHS)

Background

Organ transplantation is presently often the only available option to repair a damaged heart. As heart donors are scarce, engineering of cardiac grafts from autologous skeletal myoblasts is a promising novel therapeutic strategy. The functionality of skeletal muscle cells in the heart milieu is, however, limited because of their inability to integrate electrically and mechanically into the myocardium. Therefore, in pursuit of improved cardiac integration of skeletal muscle grafts we sought to modify primary skeletal myoblasts by overexpression of the main gap-junctional protein connexin 43 and to study electrical coupling of connexin 43 overexpressing myoblasts to cardiac myocytes in vitro.

Methods

To create an efficient means for overexpression of connexin 43 in skeletal myoblasts we constructed a bicistronic retroviral vector MLV-CX43-EGFP expressing the human connexin 43 cDNA and the marker EGFP gene. This vector was employed to transduce primary rat skeletal myoblasts in optimised conditions involving a concomitant use of the retrovirus immobilising protein RetroNectin^® and the polycation transduction enhancer Transfectam^®. The EGFP-positive transduced cells were then enriched by flow cytometry.

Results

More than four-fold overexpression of connexin 43 in the transduced skeletal myoblasts, compared with non-transduced cells, was shown by Western blotting. Functionality of the overexpressed connexin 43 was demonstrated by microinjection of a fluorescent dye showing enhanced gap-junctional intercellular transfer in connexin 43 transduced myoblasts compared with transfer in non-transduced myoblasts. Rat cardiac myocytes were cultured in multielectrode array culture dishes together with connexin 43/EGFP transduced skeletal myoblasts, control non-transduced skeletal myoblasts or alone. Extracellular field action potential activation rates in the co-cultures of connexin 43 transduced skeletal myoblasts with cardiac myocytes were significantly higher than in the co-cultures of non-transduced skeletal myoblasts with cardiac myocytes and similar to the rates in pure cultures of cardiac myocytes.

Conclusion

The observed elevated field action potential activation rate in the co-cultures of cardiac myocytes with connexin 43 transduced skeletal myoblasts indicates enhanced cell-to-cell electrical coupling due to overexpression of connexin 43 in skeletal myoblasts. This study suggests that retroviral connexin 43 transduction can be employed to augment engineering of the electrocompetent cardiac grafts from patients' own skeletal myoblasts.     

Conventional kinesin holoenzymes are composed of heavy and light chain homodimers

Conventional kinesin is a major microtubule-based motor protein responsible for anterograde transport of various membrane-bounded organelles (MBO) along axons. Structurally, this molecular motor protein is a tetrameric complex composed of two heavy (kinesin-1) chains and two light chain (KLC) subunits. The products of three kinesin-1 (kinesin-1A, -1B, and -1C, formerly KIF5A, -B, and -C) and two KLC (KLC1, KLC2) genes are expressed in mammalian nervous tissue, but the functional significance of this subunit heterogeneity remains unknown. In this work, we examine all possible combinations among conventional kinesin subunits in brain tissue. In sharp contrast with previous reports, immunoprecipitation experiments here demonstrate that conventional kinesin holoenzymes are formed of kinesin-1 homodimers. Similar experiments confirmed previous findings of KLC homodimerization. Additionally, no specificity was found in the interaction between kinesin-1s and KLCs, suggesting the existence of six variant forms of conventional kinesin, as defined by their gene product composition. Subcellular fractionation studies indicate that such variants associate with biochemically different MBOs and further suggest a role of kinesin-1s in the targeting of conventional kinesin holoenzymes to specific MBO cargoes. Taken together, our data address the combination of subunits that characterize endogenous conventional kinesin. Findings on the composition and subunit organization of conventional kinesin as described here provide a molecular basis for the regulation of axonal transport and delivery of selected MBOs to discrete subcellular locations.