The down-regulation of clusterin expression enhances the αsynuclein aggregation process

Parkinson’s Disease (PD) is a progressive neurodegenerative disease characterized by the presence of proteinaceous aggregates of αSynuclein (αSyn) in the dopaminergic neurons. Chaperones are key components of the proteostasis network that are able to counteract αSyn’s aggregation, as well as its toxic effects. Clusterin (CLU), a molecular chaperone, was consistently found to interfere with Aβ aggregation in Alzheimer’s Disease (AD). However, its role in PD pathogenesis has yet to be extensively investigated. In this study, we assessed the involvement of CLU in the αSyn aggregation process by using SH-SY5Y cells stably overexpressing αSyn (SH-Syn). First, we showed that αSyn overexpression caused a strong increase in CLU expression without affecting levels of Hsp27, Hsp70, and Hsp90, which are the chaperones widely recognized to counteract αSyn burden. Then, we demonstrated that αSyn aggregation, induced by proteasome inhibition, determines a strong increase of CLU in insoluble aggregates. Remarkably, we revealed that CLU down-regulation results in an increase of αSyn aggregates in SH-Syn without significantly affecting cell viability and the Unfolded Protein Response (UPR). Furthermore, we demonstrated the direct molecular interaction between CLU and αSyn via a co-immunoprecipitation (co-IP) assay. All together, these findings provide incontrovertible evidence that CLU is an important player in the response orchestrated by the cell to cope with αSyn burden

A Novel Gene Signature for Molecular Diagnosis of Human Prostate Cancer by RT-qPCR

Prostate cancer (CaP) is one of the most relevant causes of cancer death in Western Countries. Although detection of CaP at early curable stage is highly desirable, actual screening methods present limitations and new molecular approaches are needed. Gene expression analysis increases our knowledge about the biology of CaP and may render novel molecular tools, but the identification of accurate biomarkers for reliable molecular diagnosis is a real challenge. We describe here the diagnostic power of a novel 8-genes signature: ornithine decarboxylase (ODC), ornithine decarboxylase antizyme (OAZ), adenosylmethionine decarboxylase (AdoMetDC), spermidine/spermine N(1)-acetyltransferase (SSAT), histone H3 (H3), growth arrest specific gene (GAS1), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and Clusterin (CLU) in tumour detection/classification of human CaP. METHODOLOGY/PRINCIPAL FINDINGS: The 8-gene signature was detected by retrotranscription real-time quantitative PCR (RT-qPCR) in frozen prostate surgical specimens obtained from 41 patients diagnosed with CaP and recommended to undergo radical prostatectomy (RP). No therapy was given to patients at any time before RP. The bio-bank used for the study consisted of 66 specimens: 44 were benign-CaP paired from the same patient. Thirty-five were classified as benign and 31 as CaP after final pathological examination. Only molecular data were used for classification of specimens. The Nearest Neighbour (NN) classifier was used in order to discriminate CaP from benign tissue. Validation of final results was obtained with 10-fold cross-validation procedure. CaP versus benign specimens were discriminated with (80+/-5)% accuracy, (81+/-6)% sensitivity and (78+/-7)% specificity. The method also correctly classified 71% of patients with Gleason score<7 versus > or =7, an important predictor of final outcome. CONCLUSIONS/SIGNIFICANCE: The method showed high sensitivity in a collection of specimens in which a significant portion of the total (13/31, equal to 42%) was considered CaP on the basis of having less than 15% of cancer cells. This result supports the notion of the "cancer field effect", in which transformed cells extend beyond morphologically evident tumour. The molecular diagnosis method here described is objective and less subjected to human error. Although further confirmations are needed, this method poses the potential to enhance conventional diagnosis

A Novel Approach for an Integrated Straw tube-Microstrip Detector

We report on a novel concept of silicon microstrips and straw tubes detector, where integration is accomplished by a straw module with straws not subjected to mechanical tension in a Rohacell $^{\circledR}$ lattice and carbon fiber reinforced plastic shell. Results on mechanical and test beam performances are reported on as well.Comment: Accepted by Transactions on Nuclear Science (2005). 11 pages, 9 figures, uses lnfprep.st

Direct evidence that late Neanderthal occupation precedes a technological shift in southwestern Italy

Objectives During the middle-to-upper Paleolithic transition (50,000 and 40,000 years ago), interaction between Neanderthals and Homo sapiens varied across Europe. In southern Italy, the association between Homo sapiens fossils and non-Mousterian material culture, as well as the mode and tempo of Neanderthal demise, are still vividly debated. In this research, we focus on the study of two human teeth by using 3D geometric morphometric approaches for a reliable taxonomical attribution as well as obtaining new radiometric dates on the archeological sequence. Material and Methods This work presents two lower deciduous molars uncovered at Roccia San Sebastiano (Mondragone-Caserta, Italy), stratigraphically associated with Mousterian (RSS1) and Uluzzian (RSS2) artifacts. To obtain a probabilistic attribution of the two RSS teeth to each reference taxa group composed of Neanderthals and Homo sapiens, we performed and compared the performance of three supervised learning algorithms (flexible discriminant analysis, multiadaptive regression splines, and random forest) on both crown and cervical outlines obtained by virtual morphometric methods. Results We show that RSS1, whose Mousterian context appears more recent than 44,800-44,230 cal BP, can be attributed to a Neanderthal, while RSS2, found in an Uluzzian context that we dated to 42,640-42,380 cal BP, is attributed to Homo sapiens. Discussion This site yields the most recent direct evidence for a Neanderthal presence in southern Italy and confirms a later shift to upper Paleolithic technology in southwestern Italy compared to the earliest Uluzzian evidence at Grotta del Cavallo (Puglia, Italy)

High-accuracy methodology for the integrative restoration of archaeological teeth by using reverse engineering techniques and rapid prototyping

The reconstruction of the original morphology of bones and teeth after sampling for physicochemical (e.g., radiocarbon and uranium series dating, stable isotope analysis, paleohistology, trace element analysis) and biomolecular analyses (e.g., ancient DNA, paleoproteomics) is appropriate in many contexts and compulsory when dealing with fossil human remains. The reconstruction protocols available to date are mostly based on manual re-integration of removed portions and can lead to an imprecise recovery of the original morphology. In this work, to restore the original external morphology of sampled teeth we used computed microtomography (microCT), reverse engineering (RE), computer-aided design (CAD) and rapid prototyping (RP) techniques to fabricate customized missing parts. The protocol was tested by performing the reconstruction of two Upper Palaeolithic human teeth from the archaeological excavations of Roccia San Sebastiano (Mondragone, Caserta, southern Italy) and Riparo I of Grotte Verdi di Pradis (Clauzetto, Pordenone, north-eastern Italy) (RSS2 and Pradis 1, respectively), which were sampled for physicochemical and biomolecular analyses. It involved a composite procedure consisting in: a) the microCT scanning of the original specimens; b) sampling; c) the microCT scanning of the specimens after sampling; d) the reconstruction of the digital 3D surfaces of the specimens before and after sampling; e) the creation of digital models of the missing/sampled portions by subtracting the 3D images of the preserved portions (after the sampling) from the images of the intact specimens (before the sampling) by using reverse engineering techniques; f) the prototyping of the missing/sampled portions to be integrated; g) the painting and application of the prototypes through the use of compatible and reversible adhesives. By following the proposed protocol, in addition to the fabrication of a physical element which is faithful to the original, it was possible to obtain a remarkable correspondence between the contact surfaces of the two portions (the original and the reconstructed one) without having to resort to any manipulation/adaptation of either element

Direct evidence that late Neanderthal occupation precedes a technological shift in southwestern Italy

Objectives: During the middle-to-upper Paleolithic transition (50,000 and 40,000 years ago), interaction between Neanderthals and Homo sapiens varied across Europe. In southern Italy, the association between Homo sapiens fossils and non-Mousterian material culture, as well as the mode and tempo of Neanderthal demise, are still vividly debated. In this research, we focus on the study of two human teeth by using 3D geometric morphometric approaches for a reliable taxonomical attribution as well as obtaining new radiometric dates on the archeological sequence. Material and Methods: This work presents two lower deciduous molars uncovered at Roccia San Sebastiano (Mondragone-Caserta, Italy), stratigraphically associated with Mousterian (RSS1) and Uluzzian (RSS2) artifacts. To obtain a probabilistic attribution of the two RSS teeth to each reference taxa group composed of Neanderthals and Homo sapiens, we performed and compared the performance of three supervised learning algorithms (flexible discriminant analysis, multiadaptive regression splines, and random forest) on both crown and cervical outlines obtained by virtual morphometric methods. Results: We show that RSS1, whose Mousterian context appears more recent than 44,800–44,230 cal BP, can be attributed to a Neanderthal, while RSS2, found in an Uluzzian context that we dated to 42,640–42,380 cal BP, is attributed to Homo sapiens. Discussion: This site yields the most recent direct evidence for a Neanderthal presence in southern Italy and confirms a later shift to upper Paleolithic technology in southwestern Italy compared to the earliest Uluzzian evidence at Grotta del Cavallo (Puglia, Italy)

The talar morphology of a hypochondroplasic dwarf: A case study from the Italian Late Antique period

This project aims to test whether geometric morphometric (GM) and trabecular analyses may be useful tools in identifying talar characteristics related to hypochondroplasia. We quantified the external and internal talar morphology of a hypochondroplasic dwarf (T17) from Modena (northern Italy) dated to the sixth century AD. External talar morphology of T17 was compared with a broad sample of modern human tali (n = 159) using GM methods. Additionally, a subsample of these tali (n = 41) was used to investigate whole talar trabecular changes in T17. Our results show that GM and trabecular analyses identify a combination of traits linked to the dwarfing disorder of hypochondroplasia. These traits include decreased scaled talar dimensions compared with normal-sized individuals, presence of an accessory antero-lateral talar facet, high bone volume fraction, and high anisotropy values throughout the entire talus. In our case study, hypochondroplasia does not appear to substantially modify external talar morphology probably due to the fast growth of the talus. We suggest that small talar dimensions are associated with hypochondroplasia. An antero-lateral talar facet may result from the talus and calcaneus coalition (i.e., talocalcaneal abnormal bridging) possibly related to an everted foot posture that was limited by overgrowth of the fibula. We suggest that high talar trabecular density and strut orientation provide insights into pathological development of the trabecular plates in T17. Finally, our study suggests that high talar trabecular density and strut orientation, and small talar dimensions, may be added as possible concomitant talar hallmarks for hypochondroplasia

Gait patterns in Prader-Willi and Down syndrome patients

<p>Abstract</p> <p>Background</p> <p>Prader-Willi (PWS) and Down Syndrome (DS) are two genetic disorders characterised by some common clinical and functional features. A quantitative description and comparison of their patterns would contribute to a deeper understanding of the determinants of motor disability in these two syndromes. The aim of this study was to measure gait pattern in PWS and DS in order to provide data for developing evidence-based deficit-specific or common rehabilitation strategies.</p> <p>Methods</p> <p>19 PWS patients (17.7-40 yr) and 21 DS patients (18-39 yr) were evaluated with an optoelectronic system and force platforms for measuring kinematic and kinetic parameters during walking. The results were compared with those obtained in a group of normal-weight controls (Control Group: CG; 33.4 + 9.6 yr).</p> <p>Results and Discussion</p> <p>The results show that PWS and DS are characterised by different gait strategies. Spatio-temporal parameters indicated a cautious, abnormal gait in both groups, but DS walked with a less stable strategy than PWS. As for kinematics, DS showed a significantly reduced hip and knee flexion, especially at initial contact and ankle range of motion than PWS. DS were characterised by lower ranges of motion (p < 0.05) in all joints than CG and PWS. As for ankle kinetics, both PWS and DS showed a significantly lower push-off during terminal stance than CG, with DS yielding the lowest values. Stiffness at hip and ankle level was increased in DS. PWS showed hip stiffness values close to normal. At ankle level, stiffness was significantly decreased in both groups.</p> <p>Conclusions</p> <p>Our data show that DS walk with a less physiological gait pattern than PWS. Based on our results, PWS and DS patients need targeted rehabilitation and exercise prescription. Common to both groups is the aim to improve hypotonia, muscle strength and motor control during gait. In DS, improving pelvis and hip range of motion should represent a major specific goal to optimize gait pattern.</p

Two- and Three-Dimensional Reconstruction and Analysis of the Straw Tubes Tomography in the Btev Experiment

A check of the eccentricity of the aluminised kapton straw tubes used in the BTeV experiment is accomplished using X-ray tomography of the section of tubes modules. 2 and 3-dimensional images of the single tubes and of the modules are reconstructed and analysed. Preliminary results show that a precision better than 40 $\mu$m can be reached on the measurement of the straw radii.Comment: Presented by F.Massa at ICATPP05, Villa Olmo (Como) Italy, November 2005. 4 pages, 8 figures, uses lnfprep.st