Frontal Lobe Contusion in Mice Chronically Impairs Prefrontal-Dependent Behavior

Traumatic brain injury (TBI) is a major cause of chronic disability in the world. Moderate to severe TBI often results in damage to the frontal lobe region and leads to cognitive, emotional, and social behavioral sequelae that negatively affect quality of life. More specifically, TBI patients often develop persistent deficits in social behavior, anxiety, and executive functions such as attention, mental flexibility, and task switching. These deficits are intrinsically associated with prefrontal cortex (PFC) functionality. Currently, there is a lack of analogous, behaviorally characterized TBI models for investigating frontal lobe injuries despite the prevalence of focal contusions to the frontal lobe in TBI patients. We used the controlled cortical impact (CCI) model in mice to generate a frontal lobe contusion and studied behavioral changes associated with PFC function. We found that unilateral frontal lobe contusion in mice produced long-term impairments to social recognition and reversal learning while having only a minor effect on anxiety and completely sparing rule shifting and hippocampal-dependent behavior.     

Alkaline phosphatase in Hirudo medicinalis L.: partial purification and study of certain molecular and catalytic properties

The authors carried out extraction and purification of an alkaline phosphatase (AP) from Hirudo medicinalis and studied certain characteristics of the enzyme. After homogenizing specimens of H. medicinalis and centrifuging the homogenate at 70,000 g, the enzyme, likely membrane-bound, was obtained in a soluble form by treating the sediment with n-butanol. It was then purified by acetone fractionation and Sephadex G-200 chromatography. A fairly good purification degree was achieved; the enzyme appears to be in a single form and displays a molecular weight of about 268,000. The optimum pH (9.5) is lower than the ones generally observed as regards APs from both Invertebrata and Mammalia. The studied AP displays a strong substrate inhibition, similar to that concerning Metazoa at a higher evolutionary level (Mollusca, Echinodermata). On the contrary, the enzyme-substrate affinity, as shown by Km value (2.447 mM), is lower than as regards APs from more advanced organisms (Echinodermata, Mammalia).     

Purification and characterization of four different alkaline phosphatases from Spirographis spallanzanii

Alkaline phosphatases (APs) present in Spirographis spallanzanii were extracted by water homogenization of butanol treatment. The enzymes solubilized as above were purified by acetone fractionation and then by DEAE-cellulose and Sephadex G-200 chromatography; separation of four AP forms was achieved, which were characterized by studying their molecular and catalytic properties. The various APs differ in molecular weight, electrophoretic mobility, optimum pH, but show similar Km values and substrate inhibition pattern. Kinetic studies carried out with several inhibitors show, in particular, the existence in these APs of a second binding site and suggest a possible role for them in the metabolism of phosphoric esters of the sugars.     

Purine Metabolism in B-Cell Lymphocytic Leukemia: A Microarray Approach

B-cell chronic lymphocytic leukemia (B-CLL) is an adult-onset highly heterogeneous malignancy characterized by a cells resistance to apoptosis rather than to highly proliferative cells. In previous research, we evidenced an imbalance of purine metabolism in B-CLL cells. Since the extracellular adenosine has been proved to induce apoptosis via A2b receptor, enzymes involved in adenosine metabolism could play an important role in apoptosis resistance of B-CLL cells. We prepared a microarray chip for the analysis of 50 selected genes that could be of interest in B-CLL: enzymes of purine de-novo, salvage and catabolic pathway, oxidative stress enzymes, and apoptotis-related proteins. Preliminary results identify many genes of purine metabolism that exhibit low or high expression, while genes involved in signal transduction and apoptosis exhibit lower alterations even if of remarkable interest. This application of microarray technique seems promising and at least a subset of these genes will be valid candidates for further studies.     

Blm10 facilitates nuclear import of proteasome core particles

Short‐lived proteins are degraded by proteasome complexes, which contain a proteolytic core particle (CP) but differ in the number of regulatory particles (RPs) and activators. A recently described member of conserved proteasome activators is Blm10. Blm10 contains 32 HEAT‐like modules and is structurally related to the nuclear import receptor importin/karyopherin β. In proliferating yeast, RP‐CP assemblies are primarily nuclear and promote cell division. During quiescence, RP‐CP assemblies dissociate and CP and RP are sequestered into motile cytosolic proteasome storage granuli (PSG). Here, we show that CP sequestration into PSG depends on Blm10, whereas RP sequestration into PSG is independent of Blm10. PSG rapidly clear upon the resumption of cell proliferation and proteasomes are relocated into the nucleus. Thereby, Blm10 facilitates nuclear import of CP. Blm10‐bound CP serves as an import receptor–cargo complex, as Blm10 mediates the interaction with FG‐rich nucleoporins and is dissociated from the CP by Ran‐GTP. Thus, Blm10 represents the first CP‐dedicated nuclear import receptor in yeast.     

PrP mRNA and protein expression in brain and PrPc in CSF in Creutzfeldt-Jakob disease MM1 and VV2

Creutzfeldt-Jakob disease (CJD) is a heterogenic neurodegenerative disorder associated with abnormal post-translational processing of cellular prion protein (PrP^c). CJD displays distinctive clinical and pathological features which correlate with the genotype at the codon 129 (methionine or valine: M or V respectively) in the prion protein gene and with size of the protease-resistant core of the abnormal prion protein PrP^sc (type 1: 20/21 kDa and type 2: 19 kDa). MM1 and VV2 are the most common sporadic CJD (sCJD) subtypes. PrP mRNA expression levels in the frontal cortex and cerebellum are reduced in sCJD in a form subtype-dependent. Total PrP protein levels and PrP^sc levels in the frontal cortex and cerebellum accumulate differentially in sCJD MM1 and sCJD VV2 with no relation between PrP^sc deposition and spongiform degeneration and neuron loss, but with microgliosis, and IL6 and TNF-α response. In the CSF, reduced PrP^c, the only form present in this compartment, occurs in sCJD MM1 and VV2. PrP mRNA expression is also reduced in the frontal cortex in advanced stages of Alzheimer disease, Lewy body disease, progressive supranuclear palsy, and frontotemporal lobe degeneration, but PrP^c levels in brain varies from one disease to another. Reduced PrP^c levels in CSF correlate with PrP mRNA expression in brain, which in turn reflects severity of degeneration in sCJD.     

Novel field assays and the comparative repellency of BioUD(®) , DEET and permethrin against Amblyomma americanum

Two new field bioassay methods were developed to compare the repellent activity of BioUD^® (containing 7.75% 2‐undecanone), 98.1% DEET and 0.5% permethrin against natural populations of nymphal Amblyomma americanum (L.) (Acari: Ixodidae). In a cloth sheet assay, pieces of material measuring 41 × 58 cm, separately treated with one of the test materials or the appropriate solvent carrier, were placed at random on the ground and baited with dry ice for 1 h. Mean numbers of ticks on repellent‐treated sheets were significantly lower than on control sheets. There was no significant difference in the number of ticks collected between sheets treated with BioUD^® and those treated with DEET. However, significantly fewer ticks were found on sheets treated with BioUD^® or DEET than on permethrin‐treated sheets. In a sock test, over‐the‐calf tube socks were treated with one of the test materials or the appropriate solvent carrier. Human volunteers wore a repellent‐treated and a corresponding carrier‐treated sock on either leg and walked randomly over an area of approximately 4000 m² for 15 min. Significantly fewer ticks were collected from socks treated with BioUD^® or DEET than from socks treated with the carrier and there was no significant difference in repellency between these two agents. No difference in the mean number of ticks collected was found between permethrin‐treated and corresponding carrier‐treated socks. To examine the mechanism of repellency of BioUD^®, a four‐choice olfactometer was used to assess spatial repellency against adult A. americanum. As expected in the absence of a repellent, when all choices were represented by water‐treated filter paper, ticks were equally distributed among the choices. When one choice consisted of BioUD^®‐treated filter paper and the remaining choices of water‐treated paper, the distribution of ticks on the repellent‐treated paper was significantly lower than might be expected to occur by chance, suggesting that repellency is at least partly achieved by an olfactory mechanism.     

First report of the repellency of 2-tridecanone against ticks

2‐Tridecanone and 2‐undecanone are both found naturally in the trichomes of wild tomato plants and are important in plant resistance to herbivory. 2‐Undecanone is the U.S. Environmental Protection Agency (EPA)‐registered active ingredient in the commercially available arthropod repellent, BioUD^®. The goal of this study was to examine the tick repellency of 2‐tridecanone. Two‐choice bioassays were conducted using 8% 2‐tridecanone vs. the repellent carrier (absolute ethanol) and compared with two‐choice studies using 8% 2‐undecanone vs. absolute ethanol. Unfed, host‐seeking adult (mixed sex) Amblyomma americanum (L.) (Acari: Ixodidae) and Dermacentor variabilis Say (Acari: Ixodidae) were used to evaluate repellency and time to repellent failure at room temperature. The present study shows in filter paper assays (0.63 mg test compound/cm²) that 2‐tridecanone was 87% repellent to A. americanum at 12 h after application, but had no statistically significant repellency at 15 h and 24 h, and was 72% repellent to D. variabilis at 15 h, but had no statistically significant repellency at 24 h. By contrast, 2‐undecanone was 74% and 75% repellent to A. americanum and D. variabilis, respectively, at 2 h after application, but no statistically significant repellency was noted at 2.5 h and 3 h. In two‐choice assays on cheesecloth, 2‐tridecanone at 0.25 mg/cm² was 85% repellent to A. americanum 6 h after application, demonstrating its potential use as an arthropod repellent that can be used on clothing without the need for formulation. No statistically significant repellency was found at 9 h or 12 h. The potential use of 2‐tridecanone as a tick repellent is discussed.