Mapping the Tray of Electron Beam Melting of Ti-6Al-4V: Properties and Microstructure.

Using an electron beam melting (EBM) printing machine (Arcam A2X, Sweden), a matrix of 225 samples (15 rows and 15 columns) of Ti-6Al-4V was produced. The density of the specimens across the tray in the as-built condition was approximately 99.9% of the theoretical density of the alloy, ρT. Tensile strength, tensile elongation, and fatigue life were studied for the as-built samples. Location dependency of the mechanical properties along the build area was observed. Hot isostatic pressing (HIP) slightly increased the density to 99.99% of ρT but drastically improved the fatigue endurance and tensile elongation, probably due to the reduction in the size and the distribution of flaws. The microstructure of the as-built samples contained various defects (e.g., lack of fusion, porosity) that were not observed in the HIP-ed samples. HIP also reduced some of the location related variation in the mechanical properties values, observed in the as-printed condition.

The HIV-1 viral synapse signals human foreskin keratinocytes to secrete thymic stromal lymphopoietin facilitating HIV-1 foreskin entry.

The complexity of signal transduction resulting from the contact of human immunodeficiency virus type 1 (HIV-1)-infected cells and mucosal cells has hampered our comprehension of HIV-1 mucosal entry. Such process is driven efficiently only by viral synapse contacts, whereas cell-free HIV-1 remains poorly infectious. Using CD4+ T-cells expressing only HIV-1 envelope inoculated on human adult foreskin tissues, we designed methodologies to identify the signals transduced in foreskin keratinocytes following HIV-1-envelope-dependent viral synapse formation. We find that the viral synapse activates the MyD88-independent TLR-4-nuclear factor (NfκB) signaling pathway in keratinocytes and the subsequent secretion of cytokines including thymic stromal lymphopoietin (TSLP), a cytokine linking innate and T-helper type 2-adaptive immune responses. Moreover, the viral synapse upregulates the non-coding microRNA miR-375, known to control TSLP, and transfection of keratinocytes with anti-miR-375 blocks significantly TSLP secretion. Thus, the secretion of TSLP by keratinocytes is induced by the viral synapse in a miR-375 controlled manner. At the tissue level, these signals translate into the epidermal redistribution of Langerhans cells and formation of conjugates with T-cells, recapitulating the initial events observed in human foreskin infection by HIV-1. These results open new possibilities for designing strategies to block mucosal HIV-1 transmission, the major pathway by which HIV-1 spreads worldwide.

Calcitonin gene-related peptide inhibits human immunodeficiency type 1 transmission by Langerhans cells via an autocrine/paracrine feedback mechanism.

AIM: Peripheral neurones innervating mucosal epithelia are in direct contact with resident immune cells, including Langerhans cells (LCs). Such neurones secrete the neuropeptide calcitonin gene-related peptide (CGRP) that modulates LCs function. We recently found that CGRP strongly inhibits human immunodeficiency virus type 1 (HIV-1) transmission, by interfering with multiple steps of mucosal LC-mediated HIV-1 transfer, including increased expression of the LC-specific lectin langerin. Herein, we investigated the anti-HIV-1 mechanism of CGRP. METHODS: In the presence of CGRP, HIV-1 transfer from LCs to CD4+ T cells was tested with viral clones using either the HIV-1 co-receptor CCR5 (R5) or CXCR4 (X4). Surface expression of CCR5, CXCR4 and langerin was evaluated by flow cytometry. CGRP secretion by LCs was measured with an enzyme immunoassay. Expression of the multimeric CGRP receptor was examined by quantitative real-time RT-PCR and immuno-fluorescent microscopy. RESULTS: Calcitonin gene-related peptide decreased transfer of HIV-1 R5, but increased that of X4. These opposing effects correlated with decreased CCR5 vs. increased CXCR4 surface expression in LCs. Inhibition of HIV-1 R5 transfer by CGRP involved signal transducer and activator of transcription 4 (STAT4) activation. Both αCGRP and ßCGRP were similarly efficient in decreasing HIV-1 R5 transfer and increasing langerin expression. LCs secreted low basal levels of endogenous CGRP, which increased markedly following CGRP treatment. CGRP also increased expression of its cognate receptor in LCs. CONCLUSION: CGRP engages a positive feedback mechanism that would further enhance its anti-HIV-1 activity. This information might be relevant for the therapeutic use of CGRP as a prophylactic agent against HIV-1.

The adult penile urethra is a novel entry site for HIV-1 that preferentially targets resident urethral macrophages.

The penile urethra is routinely targeted by sexually transmitted bacterial and viral pathogens, and also represents a probable site for HIV type-1 (HIV-1) entry. Yet, the mechanisms of urethral HIV-1 transmission are unknown. To describe the initial steps of penile HIV-1 entry, we obtained whole penile tissues from individuals undergoing elective gender reassignment and developed ex vivo polarized explants of different penile epithelia, as well as in vitro immunocompetent reconstructed urethra. In penile explants, 1 h exposure to cell-associated HIV-1 results in higher HIV-1 entry into the urethra, whereas the fossa navicularis and glans are relatively resistant to HIV-1. CCR5+/CD4+ urethral macrophages are the initial cells infected by HIV-1, which exit the epithelial compartment following inoculation with cell-associated HIV-1 that induces decreased CCL2/MCP-1 production. Urethral T cells are mostly CD8+ or naive CD4+, and not infected by HIV-1 on its early entry. In urethral reconstructions, efficient translocation of cell-associated HIV-1 depends on viral tropism (R5>X4) and can be decreased by gp41-specific IgAs. Cell-free HIV-1 is inefficient at urethral penetration. Our results identify the male urethra as a novel entry site for HIV-1 that targets resident urethral macrophages. These results might explain the incomplete prophylactic efficacy of male circumcision in reducing HIV-1 transmission.

Within 1 h, HIV-1 uses viral synapses to enter efficiently the inner, but not outer, foreskin mucosa and engages Langerhans-T cell conjugates.

Although circumcision reduces male acquisition of human immunodeficiency virus type-1 (HIV-1) by 60%, the initial mechanisms of HIV-1 transmission at the foreskin remain elusive. We have established two novel and complementary models of the human adult foreskin epithelium, namely, ex vivo foreskin explants and in vitro reconstructed immunocompetent foreskins. In these models, efficient HIV-1 transmission occurs after 1 h of polarized exposure of the inner, but not outer, foreskin to mononuclear cells highly infected with HIV-1, but not to cell-free virus. HIV-1-infected cells form viral synapses with apical foreskin keratinocytes, leading to polarized budding of HIV-1, which is rapidly internalized by Langerhans cells (LCs) in the inner foreskin. In turn, LCs migrate toward the epidermis-dermis interface to form conjugates with T cells, thereby transferring HIV-1. Seminal plasma mixed with cervicovaginal secretions inhibits HIV-1 translocation. This set of results rationalizes at the cellular level the apparent protective outcome of circumcision against HIV-1 acquisition by men.

Testing system for ferromagnetic shape memory microactuators.

Ferromagnetic shape memory alloys are a class of smart materials that exhibit a unique combination of large strains and fast response when exposed to magnetic field. Accordingly, these materials have significant potential in motion generation applications such as microactuators and sensors. This article presents a novel experimental system that measures the dynamic magnetomechanical behavior of microscale ferromagnetic shape memory specimens. The system is comprised of an alternating magnetic field generator (AMFG) and a mechanical loading and sensing system. The AMFG generates a dynamic magnetic field that periodically alternates between two orthogonal directions to facilitate martensitic variant switching and to remotely achieve a full magnetic actuation cycle, without the need of mechanical resetting mechanisms. Moreover, the AMFG is designed to produce a magnetic field that inhibits 180 degrees magnetization domain switching, which causes energy loss without strain generation. The mechanical loading and sensing system maintains a constant mechanical load on the measured specimen by means of a cantilever beam, while the displacement is optically monitored with a resolution of approximately 0.1 microm. Preliminary measurements using Ni(2)MnGa single crystal specimens, with a cross section of 100x100 microm(2), verified their large actuation strains and established their potential to become a material of great importance in microactuation technology.

Antibodies to glutamate receptor subtype 3 (GluR3) are found in some patients suffering from epilepsy as the main disease, but not in patients whose epilepsy accompanies antiphospholipid syndrome or Sneddon's syndrome.

Autoantibodies (Ab's) to the "B" peptide (amino acids 372-395) of glutamate/AMPA receptor subtype 3 (GluR3) are found in serum and cerebrospinal fluid of some patients with different types of epilepsy. Since such anti-GluR3B Ab's can activate and/or kill neurons in vitro and in vivo, they may contribute to epilepsy. To investigate whether anti-GluR3B Ab's may also be relevant to epilepsy when it accompanies some autoimmune-diseases, we tested for these Ab's in patients suffering from epilepsy that accompanies anti-phospholipid syndrome (APS) or Sneddon's syndrome (SNS), both being autoimmune-diseases with frequent neurological complications. We tested 77 pediatric patients whose epilepsy is their main disease; 31 adult patients whose epilepsy accompanies APS (primary or SLE-associated) or SNS; 45 epilepsy-free APS and SNS patients; and 90 healthy controls. Compared to the controls, significantly elevated anti-GluR3B Ab's were found in 22/77 (29%) patients whose epilepsy is their main disease, but in none of the patients whose seizures accompany APS or SNS. Yet, all the APS and SNS patients harbored the characteristic anti-phospholipid Ab's (aPL), directed against cardiolipin and beta2-glycoprotein I, and had lupus anti-coagulant. Thus, anti-GluR3B Ab's are not crossreactive with aPL, and not produced as a non-specific consequence of seizures on the one hand, or autoimmune-diseases on the other. Taken together with new findings accumulated recently in our lab, we suggest that anti-GluR3B Ab's are produced primarily in the periphery due to specific/non-specific "irritation" of the immune system, and that once they reach the brain via a leaky blood-brain barrier they may cause neuronal/glial damage and facilitate the outburst of epilepsy and additional neurological abnormalities. In contrast, the presence of anti-GluR3B Ab's does not seem to increase the probability of developing APS, SNS or the seizures that often accompany these autoimmune-diseases. These findings may have important diagnostic and therapeutic implications.

Dopamine interacts directly with its D3 and D2 receptors on normal human T cells, and activates beta1 integrin function.

Dopamine by itself has not up to now been reported to activate T cell function. We show here that dopamine interacts directly with dopaminergic receptors on normal human T cells and triggers beta1 integrin-mediated T cell adhesion to a major extracellular matrix component, fibronectin (FN). Such adhesion is a characteristic feature of activated T cells, and is critical for trafficking and extravasation of T cells across blood vessels and tissue barriers. Seven dopamine D2/D3 receptor agonists and antagonists were used to identify the receptor subtypes with which dopamine specifically interacts to activate T cells. The D3 dopamine receptor agonist, 7-hydroxy-DPAT (DPAT), mimics the effects of dopamine, and the effects of both dopamine and DPAT are blocked by a specific D3 receptor antagonist, U-maleate. The dopamine receptor agonists bromocriptine and pergolide mimic the direct effect of dopamine on the beta1 integrin function, while the dopamine receptor antagonists butaclamol and haloperidol suppress it, suggesting additional signaling via the dopamine D2 receptor subtype. Our study shows, for the first time, that dopamine can directly activate T cells via ist specific receptors and suggests a possible role for dopamine in integrin-mediated cellular trafficking and extravasation of T cells in the central nervous system and possibly also in the periphery. Finally, we suggest that the reported changes in the D3 and D2 receptor RNA levels in peripheral blood lymphocytes of individuals with schizophrenia, Parkinson's disease, Alzheimer's disease and migraine can serve not only as a 'passive' diagnostic marker, but primarily reflect the dynamic functional dopamine-T cell interactions in these diseases.

Analysis of strength properties of light-cured resin composites.

OBJECTIVES: To determine and correlate the compressive and tensile strengths of resin composites, to scale their failure probability and to analyze their failure mode under combined state of stresses. METHODS: Ten brands of composites were tested for compressive and diametral tensile strengths. A recently introduced device for testing of pure shear stresses was modified to adapt to smaller specimens. Uniformity of pure shear stress distribution in the significant section was verified by a photoelastic model. Loading specimens in pure shear up to failure determined their mode of fracture under combined state of stresses. RESULTS: Diametral tensile strength yielded values that were 20% of their respective compressive strength. Multiple comparison test indicated that strength properties of the tensile strength test were much more sensitive in predicting differences between resin composites when compared to a compressive strength test. Pertac (Espe) had the highest compressive strength, Graft LC (GC) and Z-100 (3M) had the highest diametral tensile strength. No correlation was found between tensile and compressive strengths. The Weibull modulus disclosed differences in the liability of the materials to fracture. When combined state of stresses were applied through the pure shear test, failure of each specimen occurred at the principal tensile planes. SIGNIFICANCE: Compressive strength cannot predict the ability of the resin composite to withstand tensile stresses. The importance of compressive strength is limited as failure of a brittle material occurs in tension.

Radiographic features of vertically fractured, endodontically treated maxillary premolars.

OBJECTIVE: The purpose of this study was to evaluate the most frequent radiographic appearance of bony lesions associated with vertically fractured roots of endodontically treated maxillary premolars. STUDY DESIGN: The radiographic features of 102 endodontically treated teeth and their periradicular areas (51 with and 51 without vertically fractured roots) were evaluated and compared. RESULTS: The predominant appearance of the periradicular area in the teeth with vertically fractured roots was the "halo" lesion (57%); by contrast, in the non-vertically fractured roots group, a "periapical" radiolucent lesion was most frequently found (55%). Angular bone loss (14%) and periodontal radiolucency (14%) were also typical radiolucent lesions in the vertically fractured teeth. CONCLUSIONS: "Halo" lesion, perilateral radiolucency, and angular resorption of the crestal bone, combined with diffuse or defined but not corticated borders, indicated a high probability of vertical root fracture in maxillary premolars.

The effect of marginal thickness on the distortion of different impression materials.

An impression of the margins of a prepared tooth and adjacent gingival sulcus must be of sufficient thickness to withstand distortion and tearing when the impression is removed from the mouth. The purpose of this study was to compare the dimensional accuracy of Elite, Examix, and Express polyvinyl siloxanes; Permadyne polyether; and Permlastic polysulfide elastomeric impression materials. These materials were used to make impressions of a metal model that simulated prepared abutments with gingival sulci of various widths. A traveling microscope was used to measure the abutments and impression widths, and the number of defects in each impression was recorded. Between 70% and 100% of the abutment impressions with sulcular widths of 0.05 mm exhibited defects, which prevented accurate measurements in this group. Express material demonstrated a high number of defects in the 0.10 and 0.16 mm sulcular width groups. No great difference in average maximum distortion values or coefficients of variation (CV) were detected among the materials used to make impressions of abutments with sulcular width groups of 0.2 to 0.4 mm. For the sulci of 0.16 mm and less, Examix and Permiastic materials exhibited distortion and a CV comparable to the impressions of the wider sulcular groups, whereas Elite and Permadyne showed greater distortions and CVs. The differences were not statistically significant (analysis of variance) because of the larger CV among the groups. The larger coefficient of variation in the 0.1 and 0.16 mm sulcular width groups demonstrated inconsistencies in obtaining good impressions of abutments with such narrow sulcular widths.

Retrievable cemented crown options on implant-supported angled abutments: a case report.

Nonaxial implant alignment often predicates the use of cemented of screw-retained angled abutments with inherent disadvantages, including reduced retrievability on screw loosening. The purposes of this article are to review the relative merits of cemented and screw-retained angled abutments and to present a method of ensuring retrievability of cemented angled abutments.