Brain Intraventricular Injection of Amyloid-β in Zebrafish Embryo Impairs Cognition and Increases Tau Phosphorylation,Effects Reversed by Lithium

Alzheimer’s disease (AD) is a devastating neurodegenerative disorder with no effective treatment and commonly diagnosed only on late stages. Amyloid-β (Aβ) accumulation and exacerbated tau phosphorylation are molecular hallmarks of AD implicated in cognitive deficits and synaptic and neuronal loss. The Aβ and tau connection is beginning to be elucidated and attributed to interaction with different components of common signaling pathways. Recent evidences suggest that non-fibrillary Aβ forms bind to membrane receptors and modulate GSK-3β activity, which in turn phosphorylates the microtubule-associated tau protein leading to axonal disruption and toxic accumulation. Available AD animal models, ranging from rodent to invertebrates, significantly contributed to our current knowledge, but complementary platforms for mechanistic and candidate drug screenings remain critical for the identification of early stage biomarkers and potential disease-modifying therapies. Here we show that Aβ1–42 injection in the hindbrain ventricle of 24 hpf zebrafish embryos results in specific cognitive deficits and increased tau phosphorylation in GSK-3β target residues at 5dpf larvae. These effects are reversed by lithium incubation and not accompanied by apoptotic markers. We believe this may represent a straightforward platform useful to identification of cellular and molecular mechanisms of early stage AD-like symptoms and the effects of neuroactive molecules in pharmacological screenings.     

High-density genetic mapping for coffee leaf rust resistance

Coffee leaf rust caused by the fungus Hemileia vastatrix causes considerable economic losses for coffee producers. Although agrochemical products can provide sufficient disease control, the use of resistant cultivars is a safer alternative. This resistance may be constrained by one or a few genetic factors, mainly those found in material originating from interspecific hybrids. In this study, the genetic analysis of an F ₂ population consisting of 224 plants derived from a crossing of Híbrido de Timor UFV 427-15 (resistant) with Catuaí Amarelo IAC 30 (susceptible) showed that a dominant gene confers the resistance of coffee to race II of H. vastatrix. From a genetic map saturated with 25 amplified fragment length polymorphism (AFLP) markers linked to the resistance gene, we developed a high-density genetic map with six sequence-characterized amplified region (SCAR) markers delimiting a chromosomal region of 9.45 cM and flanking the dominant gene at 0.7 and 0.9 cM. This is the first saturated and high-density genetic map obtained from this region containing the resistance gene. The results of this study are of great importance for the introduction of molecular markers for marker-assisted selection; they will also facilitate studies related to the cloning, structure, and function of race-specific genes involved in the resistance of coffee trees to H. vastatrix.     

Profiling the changes in signaling pathways in ascorbic acid/β‐glycerophosphate‐induced osteoblastic differentiation

Despite numerous reports on the ability of ascorbic acid and β‐glycerophosphate (AA/β‐GP) to induce osteoblast differentiation, little is known about the molecular mechanisms involved in this phenomenon. In this work, we used a peptide array containing specific consensus sequences (potential substrates) for protein kinases and traditional biochemical techniques to examine the signaling pathways modulated during AA/β‐GP‐induced osteoblast differentiation. The kinomic profile obtained after 7 days of treatment with AA/β‐GP identified 18 kinase substrates with significantly enhanced or reduced phosphorylation. Peptide substrates for Akt, PI3K, PKC, BCR, ABL, PRKG1, PAK1, PAK2, ERK1, ERBB2, and SYK showed a considerable reduction in phosphorylation, whereas enhanced phosphorylation was observed in substrates for CHKB, CHKA, PKA, FAK, ATM, PKA, and VEGFR‐1. These findings confirm the potential usefulness of peptide microarrays for identifying kinases known to be involved in bone development in vivo and in vitro and show that this technique can be used to investigate kinases whose function in osteoblastic differentiation is poorly understood. J. Cell. Biochem. 112: 71–77, 2011. © 2010 Wiley‐Liss, Inc.     

Determination of the critical micelle concentration of surfactants using the fluorescent probe N-phenyl-1-naphthylamine

The fluorescence quantum yield of N-phenyl-1-naphthylamine (NPN) increases about 10-fold and the wavelength of maximum fluorescence emission is blue-shifted when this molecule partitions into the apolar core of micellar structures from the aqueous phase. This property allowed the utilization of NPN as a fluorescent indicator of micelle formation by 14 different surfactants belonging to the families of alkyltrimethylammonium halides, alkylsulfates, alkylbetaines, alkylglucosides, and bile salts. The critical micelle concentrations (CMCs) determined with NPN agreed well with literature values. In this work NPN was used at a concentration of 10(-6) M which allowed determination of CMCs in the range between approximately 10(-5) and greater than 10(-2) M. With high-sensitivity instrumentation considerably lower NPN concentrations can be used and consequently considerably lower CMCs can be rapidly and accurately determined.     

Unpacking of a Crumpled Wire from Two-Dimensional Cavities

The physics of tightly packed structures of a wire and other threadlike materials confined in cavities has been explored in recent years in connection with crumpled systems and a number of topics ranging from applications to DNA packing in viral capsids and surgical interventions with catheter to analogies with the electron gas at finite temperature and with theories of two-dimensional quantum gravity. When a long piece of wire is injected into two-dimensional cavities, it bends and originates in the jammed limit a series of closed structures that we call loops. In this work we study the extraction of a crumpled tightly packed wire from a circular cavity aiming to remove loops individually. The size of each removed loop, the maximum value of the force needed to unpack each loop, and the total length of the extracted wire were measured and related to an exponential growth and a mean field model consistent with the literature of crumpled wires. Scaling laws for this process are reported and the relationship between the processes of packing and unpacking of wire is commented upon.     

Prenatal haloperidol alters the expression of DNA polymerases in brain regions of neonate rats

1. Previous studies have reported a marked reduction in the [3H]thymidine incorporation in forebrain after administration of a dopamine antagonist such as haloperidol. 2. We have investigated the possibility that the expression levels of genes related to DNA metabolism could be altered by haloperidol treatment. 3. By Northern blot analysis, we have studied the steady-state mRNA levels for genes involved in DNA metabolism, in neonate rat mesencephalon and forebrain, after chronic prenatal blockade of dopamine receptors with haloperidol. 4. We found that the expression levels for DNA polymerases alpha and beta were clearly reduced in forebrain by haloperidol treatment. On the contrary, the expression of DNA polymerase beta was increased in mesencephalon. 5. Our results suggest that dopamine receptors occupancy may be a critical factor in controlling cell proliferation during brain development, through a mechanism(s) involving changes in the expression of DNA polymerases.     

Selection and characterization of mexican strains ofBacillus thuringiensis active against four major lepidopteran maize pests

In order to isolate novel delta-endotoxins fromBacillus thuringiensis Berliner, a total of 426 native isolates (in varying numbers for each pest) were screened against four major maize pests: corn earworm,Helicoverpa zea; fall armyworm,Spodoptera frugiperda; southwestern corn borer,Diatraea graridiosella, and sugarcane borer,Diatraea saccharalis. Spore-crystal complexes from the isolates were integrated into semi-artificial diets of each pest and mortality was assessed 7 days after treatment. A total of 25 isolates were selected on the basis of highest larval mortality against at least one insect species. There was no correspondence of the most toxic isolates when tested against the four different insect species. Most of the 25 selected isolates caused higher toxicities against all four pests than the standard strain HD-1, regardless of not achieving 100% mortality in any bioassay.H. zea demonstrated the highest level of mortality (96%) and was susceptible to the largest number of isolates (98). None of the other insect species were found susceptible at levels greater than 60%. All the selected active strains were isolated from stored grain dusts (except for LBIT-167), and had bipyramidal crystals with Cry I-like proteins. Most isolates also formed an associated square (cubic) inclusion, with Cry Il-like proteins according to SDS-PAGE analysis of their parasporal bodies. The most active isolates will be subjected to further studies, in order to identify putative novel genes to be expressed in transgenic maize     

The Potential Impact of White-Nose Syndrome on the Conservation Status of North American Bats

White-Nose syndrome (WNS) is an emergent infectious disease that has already killed around six million bats in North America and has spread over two thousand kilometers from its epicenter. However, only a few studies on the possible impacts of the fungus on bat hosts were conducted, particularly concerning its implications for bat conservation. We predicted the consequences of WNS spread by generating a map with potential areas for its occurrence based on environmental conditions in sites where the disease already occurs, and overlaid it with the geographic distribution of all hibernating bats in North America. We assumed that all intersection localities would negatively affect local bat populations and reassessed their conservation status based on their potential population decline. Our results suggest that WNS will not spread widely throughout North America, being mostly restricted to the east and southeast regions. In contrast, our most pessimistic scenario of population decline indicated that the disease would threaten 32% of the bat species. Our results could help further conservation plans to preserve bat diversity in North America.