Trypanosoma cruzi high infectivity in vitro is related to cardiac lesions during long-term infection in Beagle dogs

Trypanosoma cruzi is a hemoflagelate parasite associated with heart dysfunctions causing serious problems in Central and South America. Beagle dogs develop the symptoms of Chagas disease in humans, and could be an important experimental model for better understanding the immunopathogenic mechanisms involved in the chagasic infection. In the present study we investigated the relation among biological factors inherent to the parasite (trypomastigote polymorphism and in vitro infectivity) and immunoglobulin production, inflammation, and fibrosis in the heart of Beagle dogs infected with either T. cruzi Y or Berenice-78 strains. In vitro infectivity of Vero cells as well as the extension of cardiac lesions in infected Beagle was higher for Y strain when compared to Berenice-78 strain. These data suggested that in vitro infectivity assays may correlate with pathogenicity in vivo. In fact, animals infected with Y strain, which shows prevalence of slender forms and high infectivity in vitro, presented cardiomegaly, inflammation, and fibrosis in heart area. Concerning the immunoglobulin production, no statistically significant difference was observed for IgA, IgM or IgG levels among T. cruzi infected animals. However, IgA together IgM levels have shown to be a good marker for the acute phase of Chagas disease.     

K+ retention in leaf mesophyll,an overlooked component of salinity tolerance mechanism: A case study for barley

Plant salinity tolerance is a physiologically complex trait, with numerous mechanisms contributing to it. In this work,we show that the ability of leaf mesophyll to retain Kt represents an important and essentially overlooked component of a salinity tolerance mechanism. The strong positive correlation between mesophyll Ktretention ability under saline conditions(quantified by the magnitude of Na Cl-induced Kt efflux from mesophyll) and the overall salinity tolerance(relative fresh weight and/or survival or damage under salinity stress) was found while screening 46 barley(Hordeum vulgare L.) genotypes contrasting in their salinity tolerance. Genotypes with intrinsically higher leaf Ktcontent under control conditions were found to possess better Ktretention ability under salinity and, hence, overall higher tolerance. Contrary to previous reports for barley roots, Ktretention in mesophyll was not associated with an increased Ht-pumping in tolerant varieties but instead correlated negatively with this trait. These findingsare explained by the fact that increased Htextrusion may be needed to charge balance the activity and provide the driving force for the high affinity HAK/KUP Kttransporters required to restore cytosolic Kthomeostasis in salt-sensitive genotypes.     

Application of high-resolution DNA melting for genotyping and variant scanning of diploid and autotetraploid potato

The ideal marker system for tetraploid potato would be dosage-sensitive and have the ability to distinguish heterozygous genotypes with multiple haplotypes within the genomic region targeted by the marker. The objective of this study was to evaluate the utility of high-resolution DNA melting (HRM) for genotyping and polymorphism detection in diploid and tetraploid potato. Amplicon scanning, unlabelled probe, and short amplicon assays were developed for four candidate genes affecting tuber skin and flesh colour, and starch, and a marker linked to nematode resistance. Genotyping a set of 95 potato clones revealed several examples of clones with three distinct haplotypes. Combined probe and amplicon analysis identified between 29 and 44 unique genotypes for the same assays. Assays developed for four of the five target genes are suitable for marker-assisted selection in potato breeding programs. This study illustrates the use of HRM in potato genetics. Further advances in the technology and associated data analysis should make HRM a useful tool for basic and applied studies of potato.     

Effect of secondary metabolites associated with anaerobic soil conditions on ion fluxes and electrophysiology in barley roots

The effects of secondary metabolites produced by waterlogged soils on net K(+), H(+), and Ca(2+) fluxes were studied in the mature zone of roots of two barley (Hordeum vulgare) cultivars contrasting in their waterlogging (WL) tolerance using the noninvasive microelectrode ion flux measuring technique. In WL-sensitive variety 'Naso Nijo', all three lower monocarboxylic acids (formic, acetic, and propionic acids) and three phenolic acids (benzoic, 2-hydroxybenzoic, 4-hydroxybenzoic acids) caused a substantial shift toward steady K(+) efflux, accompanied by an immediate net influx of H(+). Detrimental effects of secondary metabolites on K(+) homeostasis in root cells were absent in WL-tolerant 'TX' variety. Root treatment with Mn(2+) caused only a temporary K(+) loss that returned to the initial level 10 min after treatment. Phenolic acids slightly increased Ca(2+) influx immediately after treatment, while other metabolites tested resulted in transient Ca(2+) efflux from the root. In the long-term (24 h) treatment, all metabolites tested significantly reduced K(+) uptake and the adverse effects of phenolic acids were smaller than for monocarboxylic acids and Mn(2+). Treatment with monocarboxylic acids for 24 h shifted H(+) from net efflux to net influx, while all three phenolic acids did not cause significant effects compared with the control. Based on results of pharmacological experiments and membrane potential measurements, a model explaining the effects of secondary metabolites on membrane transport activity is proposed. We also suggest that plant tolerance to these secondary metabolites could be considered a useful trait in breeding programs.     

Protodrilus (Protodrilidae, Annelida) from the southern and southeastern Brazilian coasts

Protodrilus corderoi, Protodrilus ovarium n. sp. and Protodrilus pythonius n. sp. are reported from beaches in southern and southeastern Brazil and described combining live observations with light and electron scanning microscopy studies. Protodrilus corderoi is redescribed from new collections at the type locality, and a neotype for the species is assigned since the original type material no longer exists. New information on reproductive organs, segmental adhesive glands and unpigmented ciliary receptors as well as morphometrics is provided. Protodrilus ovarium n. sp. and P. pythonius n. sp. are formally described. Protodrilus ovarium n. sp. is diagnosed by the presence of separated lateral organs on segments 7–12, three spermioducts of segments 10–12 and salivary glands in segments 1–9. Protodrilus pythonius n. sp. is defined by the presence of separated lateral organs on segments 7–16, long pygidial lobes and body tapering toward the pygidium. The distribution of the different species in more or less spacious habitats seems to be correlated with their gross morphology. Protodrilus pythonius n. sp., with relatively long and wide body and long palps with ciliary bands, was collected in very coarse sandy sediments at a reflective sheltered beach. Conversely, P. corderoi and P. ovarium n. sp., both possessing more slender bodies with shorter, less ciliated palps, occurred in medium-coarse, well-sorted sediments in the more energetic swash zone of exposed intermediate-reflective beaches. The finding of P. pythonius and P. corderoi in nearby beaches corroborates other studies showing a higher morphological variability among species in different habitats within the same geographical area than among species in the same habitat in different geographical areas.     

Enhanced tumor responses to dendritic cells in the absence of CD8-positive cells

Wild-type mice immunized with MART-1 melanoma Ag-engineered dendritic cells (DC) generate strong Ag-specific immunity that has an absolute requirement for both CD8(+) and CD4(+) T cells. DC administration to CD8 alpha knockout mice displayed unexpectedly enhanced levels of protection to tumor challenge despite this deficiency in CD8(+) T cells and the inability to mount MHC class I-restricted immune responses. This model has the following features: 1) antitumor protection is Ag independent; 2) had an absolute requirement for CD4(+) and NK1.1(+) cells; 3) CD4(+) splenocytes are responsible for cytokine production; 4) lytic cells in microcytotoxicity assays express NK, but lack T cell markers (NK1.1(+) alpha beta TCR(-) CD3(-)); and 5) the lytic phenotype can be transferred to naive CD8 alpha knockout mice by NK1.1(+) splenocytes. Elucidation of the signaling events that activate these effective cytotoxic cells and the putative suppressive mechanisms in a wild-type environment may provide means to enhance the clinical activity of DC-based approaches.     

In vivo analysis of protein kinase B (PKB)/Akt regulation in DNA-PKcs-null mice reveals a role for PKB/Akt in DNA damage response and tumorigenesis

Full activation of protein kinase B (PKB/Akt) requires phosphorylation on Thr-308 and Ser-473. It is well established that Thr-308 is phosphorylated by 3-phosphoinositide-dependent kinase-1 (PDK1). Ser-473 phosphorylation is mediated by both mammalian target of rapamycin-rictor complex (mTORC2) and DNA-dependent protein kinase (DNA-PK) depending on type of stimulus. However, the physiological role of DNA-PK in the regulation of PKB phosphorylation remains to be established. To address this, we analyzed basal, insulin-induced, and DNA damage-induced PKB Ser-473 phosphorylation in DNA-PK catalytic subunit-null DNA-PKcs(-/-) mice. Our results revealed that DNA-PK is required for DNA damage-induced phosphorylation but dispensable for insulin- and growth factor-induced PKB Ser-473 phosphorylation. Moreover, DNA-PKcs(-/-) mice showed a tissue-specific increase in basal PKB phosphorylation. In particular, persistent PKB hyperactivity in the thymus apparently contributed to spontaneous lymphomagenesis in DNA-PKcs(-/-) mice. Significantly, these tumors could be prevented by deletion of PKBalpha. These findings reveal stimulus-specific regulation of PKB activation by specific upstream kinases and provide genetic evidence of PKB deregulation in DNA-PKcs(-/-) mice.     

GranatumX: A Community-engaging,Modularized, and Flexible Webtool for Single-cell Data Analysis

We present GranatumX, a next-generation software environment for single-cell RNA sequencing (scRNA-seq) data analysis. GranatumX is inspired by the interactive webtool Granatum. GranatumX enables biologists to access the latest scRNA-seq bioinformatics methods in a web-based graphical environment. It also offers software developers the opportunity to rapidly promote their own tools with others in customizable pipelines. The architecture of GranatumX allows for easy inclusion of plugin modules, named Gboxes, which wrap around bioinformatics tools written in various programming languages and on various platforms. GranatumX can be run on the cloud or private servers and generate reproducible results. It is a community-engaging, flexible, and evolving software ecosystem for scRNA-seq analysis, connecting developers with bench scientists. GranatumX is freely accessible at http://garmiregroup.org/granatumx/app.     

Physical fitness, menstrual cycle disorders and smoking habit in Croatian National Ballet and National Folk Dance Ensembles

The study investigated differences in morphological, motor and functional abilities between folk and ballet dancers. The sample comprised 51 female subjects: Croatian National Ballet (N=30) and Croatian National Folk Ensemble "LADO" (N=21). The data regarding menstrual cycle, menarche, number of births and smoking habit were collected and the morphological, motor and functional abilities measured. Significant correlations between the amount of fat tissue and number of births were found in both groups. Folk dancers were as tall as ballet dancers but weighted more and had a larger body frame (p<0.001). Ballet dancers were more flexible but there were no differences in absolute maximal oxygen uptake (2.65 vs. 2.35 L/min, p=0.101). Still, as the ballet dancers weighted less, their relative maximal oxygen uptake was significantly higher (37.62 vs. 50.22 mL/kg/min, p<0.001). Also, a high number of 45% of smokers among professional ballet and professional folk dancers was found.