Etk/Bmx Regulates Proteinase-Activated-Receptor1 (PAR1) in Breast Cancer Invasion: Signaling Partners,Hierarchy and Physiological Significance

Background

While protease-activated-receptor 1 (PAR₁) plays a central role in tumor progression, little is known about the cell signaling involved.

Methodology/Principal Findings

We show here the impact of PAR₁ cellular activities using both an orthotopic mouse mammary xenograft and a colorectal-liver metastasis model in vivo, with biochemical analyses in vitro. Large and highly vascularized tumors were generated by cells over-expressing wt hPar1, Y397Z hPar1, with persistent signaling, or Y381A hPar1 mutant constructs. In contrast, cells over-expressing the truncated form of hPar1, which lacks the cytoplasmic tail, developed small or no tumors, similar to cells expressing empty vector or control untreated cells. Antibody array membranes revealed essential hPar1 partners including Etk/Bmx and Shc. PAR₁ activation induces Etk/Bmx and Shc binding to the receptor C-tail to form a complex. Y/A mutations in the PAR₁ C-tail did not prevent Shc-PAR₁ association, but enhanced the number of liver metastases compared with the already increased metastases obtained with wt hPar1. We found that Etk/Bmx first binds via the PH domain to a region of seven residues, located between C378-S384 in PAR₁ C-tail, enabling subsequent Shc association. Importantly, expression of the hPar1-7A mutant form (substituted A, residues 378-384), which is incapable of binding Etk/Bmx, resulted in inhibition of invasion through Matrigel-coated membranes. Similarly, knocking down Etk/Bmx inhibited PAR₁-induced MDA-MB-435 cell migration. In addition, intact spheroid morphogenesis of MCF10A cells is markedly disrupted by the ectopic expression of wt hPar1. In contrast, the forced expression of the hPar1-7A mutant results in normal ball-shaped spheroids. Thus, by preventing binding of Etk/Bmx to PAR₁ -C-tail, hPar1 oncogenic properties are abrogated.

Conclusions/Significance

This is the first demonstration that a cytoplasmic portion of the PAR₁ C-tail functions as a scaffold site. We identify here essential signaling partners, determine the hierarchy of binding and provide a platform for therapeutic vehicles via definition of the critical PAR₁ -associating region in the breast cancer signaling niche.     

On the significance of genital characters in supraspecific systematics of the elaterid subfamily agrypninae (Coleoptera,Elateridae)

The genital characters of some click beetles from the tribes Agrypnini and Conoderini of the subfamily Agrypninae are reviewed. The structural features of male sternite IX, aedeagus, and bursa copulatrix are shown to have great diagnostic value for supraspecific taxa. Based on these characters, the distinctness of the genus Compsolacon Rtt. is confirmed and the taxonomic position of some species from the tribe Conoderini is clarified. Agrypnus cordicollis (Cand.) is recorded for the Russian fauna for the first time.     

Excitability of the Motor Cortex in De Novo Patients with Celiac Disease

Introduction

Celiac disease (CD) may initially present as a neurological disorder or may be complicated by neurological changes. To date, neurophysiological studies aiming to an objective evaluation of the potential central nervous system involvement in CD are lacking.

Objective

To assess the profile of cortical excitability to Transcranial Magnetic Stimulation (TMS) in a group of de novo CD patients.

Materials and methods

Twenty CD patients underwent a screening for cognitive and neuropsychiatric symptoms by means of the Mini Mental State Examination and the Structured Clinical Interview for DSM-IV Axis I Disorders, respectively. Instrumental exams, including electroencephalography and brain computed tomography, were also performed. Cortico-spinal excitability was assessed by means of single and paired-pulse TMS using the first dorsal interosseus muscle of the dominant hand. TMS measures consisted of resting motor threshold, motor evoked potentials, cortical silent period (CSP), intracortical inhibition (ICI) and facilitation (ICF). None of the CD was on gluten-free diet. A group of 20 age-matched healthy controls was used for comparisons.

Results

CD showed a significantly shorter CSP (78.0 vs 125.0 ms, p<0.025), a reduced ICI (0.3 vs 0.2, p<0.045) and an enhanced ICF (1.1 vs 0.7, p<0.042) compared to controls. A dysthymic disorder was identified in five patients. The effect size between dysthymic and non-dysthymic CD patients indicated a low probability of interference with the CSP (Cohen''s d -0.414), ICI (-0.278) and ICF (-0.292) measurements.

Conclusion

A pattern of cortical excitability characterized by “disinhibition” and “hyperfacilitation” was found in CD patients. Immune system dysregulation might play a central role in triggering changes of the motor cortex excitability.     

Prevalence of Trachoma in the North Region of Cameroon: Results of a Survey in 15 Health Districts

Background

To estimate the prevalence of trachoma in the North Region of Cameroon in order to facilitate the planning of trachoma control activities in this region, a survey was carried out in 2011 and 2012 in 15 health districts (HDs).

Methodology

A cross-sectional, two-stage cluster random sampling survey was carried out. The survey focused on two target populations: children aged 1 to 9 years for the prevalence of Trachomatous Inflammation-Follicular (TF) and those aged 15 and over for the prevalence of Trachomatous Trichiasis (TT). The sample frame was an exhaustive list of villages and neighborhoods of HDs. The World Health Organization simplified trachoma grading system was used for the recognition and registration of cases of trachoma.

Principal Findings

30,562 children aged 1 to 9 years and 24,864 people aged 15 and above were examined. In children aged 1–9 years, the overall prevalence of TF was 4.2% (95% confidence intervals (CI): 4.0–4.5%). Three (3) of 15 HDs in the region showed TF prevalence of ≥10% (Poli, Rey Bouba, and Tcholliré). The overall TT prevalence was 0.25% (95% CI: 0.20–0.33%). There were estimated 1265 TT cases in the region. The prevalence of blindness was 0.01% (95% CI: 0.00–0.03%), low vision was 0.11% (95% CI: 0.07–0.17%), and corneal opacity was 0.22% (95% CI: 0.17–0.29%).

Conclusions/Significance

This survey provides baseline data for the planning of activities to control trachoma in the region. The overall prevalence of TF in the region is 4.2%, and that of TT is 0.2%; three HDs have a TF prevalence ≥10%. These three HDs are eligible for mass drug administration with azythromycin, along with the implementation of the “F” and “E” components of the SAFE strategy.     

The role of monocyte/macrophages as vehicles of dissemination of Simkania negevensis: an in vitro simulation model

Exposure to Simkania negevensis (Sn), an intracellular microorganism that has been associated with respiratory tract infections in infants and adults, is prevalent. Sn can multiply within free-living amoebae and has been detected in domestic water supplies, which may constitute a source of infection with the organism. Its path of transport from its portal of entry to the body to its target organs is unknown. In this study, the possibility that monocytes/macrophages may serve as vehicles of transmission was examined. In vitro cocultivation of Sn-infected Acanthamoeba polyphaga with the monocyte/macrophage cell line U937 resulted in the death of the amoebae and infection of the U937 cells. Sn entered and multiplied in U937 cells within short periods of time, and the microorganism could be transferred from U937 cells to cell cultures of various origins. Uninfected monocyte/macrophages could become infected when in contact with either actively or persistently Sn-infected cell cultures. Persistently infected cultures in contact with uninfected U937 cells became actively infected. The results of this study provide a basis for determination of the molecular mechanisms of monocyte/macrophage-cell interactions in transfer of infection and may contribute to a better understanding of the pathogenesis of Sn infections in vivo.