Non-paraxial design and fabrication of a compact OAM sorter in the telecom infrared.

A novel optical device is designed and fabricated in order to overcome the limits of the traditional sorter based on log-pol optical transformation for the demultiplexing of optical beams carrying orbital angular momentum (OAM). The proposed configuration simplifies the alignment procedure and significantly improves the compactness and miniaturization level of the optical architecture. Since the device requires to operate beyond the paraxial approximation, a rigorous formulation of transformation optics in the non-paraxial regime has been developed and applied. The sample has been fabricated as 256-level phase-only diffractive optics with high-resolution electron-beam lithography, and tested for the demultiplexing of OAM beams at the telecom wavelength of 1310 nm. The designed sorter can find promising applications in next-generation optical platforms for mode-division multiplexing based on OAM modes both for free-space and multi-mode fiber transmission.

Induction of anti-tumor CD8 T cell responses by experimental ECP-induced human dendritic antigen presenting cells.

Extracorporeal photochemotherapy (ECP), or photopheresis, is distinguished by the specificity of the clinically potent immunologic reactions it initiates or regulates. The selectivity of ECP-induced immunoprotection for the malignant clone in cutaneous T cell lymphoma (CTCL), and for the pathogenic clones in allograft rejection and graft-versus-host disease (GVHD), has suggested a central mechanistic role for dendritic antigen presenting cells (DC). Discovery of ECP's induction of monocyte-derived DC, via monocyte signaling by ECP-plate activated platelets, and the absolute dependency of experimental ECP on such induced DC, supports that premise. Herein, we show that ECP-induced DC are capable of stimulating CD8 T cell responses to tumor antigens with which they are loaded. They internalize an antigen-specific melanoma-associated protein then present it onto a class I major histocompatibility, which then stimulates expansion of anti-tumor CD8 T cell populations. We conclude that ECP-induced DC prominently contribute to its initiation of anti-tumor immunity and raise the possibility that the therapy may be applicable to the immunotherapeutic management of a broader spectrum of cancers.

Integrin-driven monocyte to dendritic cell conversion in modified extracorporeal photochemotherapy.

Due to clinical efficacy and safety profile, extracorporeal photochemotherapy (ECP) is a commonly used cell treatment for patients with cutaneous T cell lymphoma (CTCL) and graft-versus-host disease (GVHD). The capacity of ECP to induce dendritic antigen-presenting cell (DC)-mediated selective immunization or immunosuppression suggests a novel mechanism involving pivotal cell signalling processes that have yet to be clearly identified as related to this procedure. In this study we employ two model systems of ECP to dissect the role of integrin signalling and adsorbed plasma proteins in monocyte-to-DC differentiation. We demonstrate that monocytes that were passed through protein-modified ECP plates adhered transiently to plasma proteins, including fibronectin, adsorbed to the plastic ECP plate and activated signalling pathways that initiate monocyte-to-DC conversion. Plasma protein adsorption facilitated 54·2 ± 4·7% differentiation, while fibronectin supported 29·8 ± 7·2% differentiation, as detected by DC phenotypic expression of membrane CD80 and CD86, as well as CD36, human leucocyte antigen D-related (HLA-DR) and cytoplasmic CD83. Further, we demonstrate the ability of fibronectin and other plasma proteins to act through cell adhesion via the ubiquitous arginine-glycine-aspartic (RGD) motif to drive monocyte-to-DC differentiation, with high-density RGD substrates supporting 54·1 ± 5·8% differentiation via αVß3 and α5ß1integrin signalling. Our results demonstrate that plasma protein binding integrins and plasma proteins operate through specific binding domains to induce monocyte-to-DC differentiation in ECP, providing a mechanism that can be harnessed to enhance ECP efficacy.

Orientational dynamics for an amphiphilic-solvent solution.

In this work, we performed Monte Carlo simulations on a lattice model for spontaneous amphiphilic aggregation, in order to study the orientational and hydrogen-bonding dynamics of water on different regions inside the micellar solution. We employed an associating lattice gas model that mimics the aqueous solvent, which presents a rich phase diagram with first- and second-order transition lines. Even though this is a simplified model, it makes possible to investigate the orientational dynamics of water in an equilibrium solution of amphiphiles, as well as the influence of the different phases of the solvent in the interfacial and bulk water dynamics. By means of extensive simulations, we showed that, at high temperatures, the behavior of the orientational relaxation and hydrogen bonding of water molecules in the bulk, first, and second hydration shells are considerable different. We observe the appearance of a very slow component for water molecules in the first hydration shell of micelles when the system reaches a high-density phase, consistent with previous theoretical and experimental studies concerning biological water. Also, at high temperatures, we find that water molecules in the second hydration shell of micelles have an orientational decay similar to that of bulk water, but with a generally slower dynamics. Otherwise, at low temperatures, we have two components for the orientational relaxation of bulk water in the low density liquid phase, and only a single component in the high density liquid (HDL) phase, which reflect the symmetry properties of the different phases of the solvent model. In the very dense region of water molecules in the first hydration shell of micelles at low temperatures, we find two components for the orientational relaxation on both liquid phases, one of them much slower than that in the single component of bulk water in the HDL phase. This happens even though our model does not present any hindrance to the water rotational freedom caused by the presence of the amphiphiles.

In vitro study of microleakage of fissure sealant with different previous treatments.

The purpose of this work was to evaluate microleakeage of a sealant after using three different techniques for conditioning the surface to be sealed. Twenty-four caries-free upper and lower premolars were used, which were preserved in distilled water at room temperature. The structural faults were enlarged using a cylindrical conical diamond (ISO 007). Teeth were randomly assigned into three groups of eight. Group I (control) was conditioned with 37% phosphoric acid (Vivadent) for 15 seconds after which the sealant Helioseal F (Vivadent) was applied and cured for 40 seconds. Group II was conditioned in the same way, after which one-step adhesive Te-econom (Vivadent) and the sealant were applied. Group III was conditioned using a self-etching adhesive, Go (SDI), after which the sealant was applied. Adhesive was applied according to the manufacturer's instructions. The samples were thermocycled for 300 cycles between 5 degrees and 55 degreesC and immersed in a 2% methylene blue solution for 48 hs. at standardized temperature of 37 degreesC +/- 1 degree. Then they were rinsed with tap water and ground longitudinally in V-P direction with silica carbide rotatory disks of decreasing grit. The amount of leakage was evaluated under stereoscopic microscope at 40X magnification. The longitudinal penetration of dye into the tooth-sealant interface was scored on a scale of 0 to 3. The results were analyzed by a Kruskal-Wallis non-parametric test. In Group II, 100% of the samples showed low (50%) or no (50%) leakage. Both the other groups had a higher percentage of specimens with high leakage (scores 2 and 3) (P = 0.000). Group II had the best performance, with significant differences (P = 0.0028) compared to the other experimental groups. Marginal leakage was lowest when the tooth was conditioned with phosphoric acid and subsequent application of an adhesive, prior to sealant.

Interplay between micelle formation and waterlike phase transitions.

A lattice model for amphiphilic aggregation in the presence of a structured waterlike solvent is studied through Monte Carlo simulations. We investigate the interplay between the micelle formation and the solvent phase transition in two different regions of temperature-density phase diagram of pure water. A second order phase transition between the gaseous (G) and high density liquid (HDL) phases that occurs at very high temperatures, and a first order phase transition between the low density liquid (LDL) and (HDL) phases that takes place at lower temperatures. In both cases, we find the aggregate size distribution curve and the critical micellar concentration as a function of the solvent density across the transitions. We show that micelle formation drives the LDL-HDL first order phase transition to lower solvent densities, while the transition G-HDL is driven to higher densities, which can be explained by the markedly different degrees of micellization in both cases. The diffusion coefficient of surfactants was also calculated in the LDL and HDL phases, changing abruptly its behavior due to the restructuring of waterlike solvent when we cross the first order LDL-HDL phase transition. To understand such behavior, we calculate the solvent density and the number of hydrogen bonds per water molecule close to micelles. The curves of the interfacial solvent density and the number of hydrogen bonds per water molecule in the first hydration signal a local phase change of the interfacial water, clarifying the diffusion mechanism of free surfactants in the solvent.

Regulation of cutaneous malignancy by gammadelta T cells.

The localization of gammadelta T cells within epithelia suggests that these cells may contribute to the down-regulation of epithelial malignancies. We report that mice lacking gammadelta cells are highly susceptible to multiple regimens of cutaneous carcinogenesis. After exposure to carcinogens, skin cells expressed Rae-1 and H60, major histocompatibility complex-related molecules structurally resembling human MICA. Each of these is a ligand for NKG2d, a receptor expressed by cytolytic T cells and natural killer (NK) cells. In vitro, skin-associated NKG2d+ gammadelta cells killed skin carcinoma cells by a mechanism that was sensitive to blocking NKG2d engagement. Thus, local T cells may use evolutionarily conserved proteins to negatively regulate malignancy.

Monte Carlo simulations for amphiphilic aggregation near a water phase transition.

In this study we analyze the equilibrium and dynamical properties of a lattice model for amphiphilic aggregation in a waterlike associating solvent. The amphiphiles are described as flexible chains of interconnected sites in a body-centered cubic lattice, with hydrophilic and hydrophobic portions. The solvent molecules occupy a single site and resemble the water tetrahedral molecular structure, with the possibility of hydrogen-bond formation and different densities. Following the phase diagram of the solvent model, we are able to study the effects of a phase transition of the solvent in the micellar dynamics. By carrying out Monte Carlo simulations, we analyze the micelle aggregate size distribution curve, the critical micelle concentration, the surfactant diffusion coefficient, the residence time, and the exit/entering rates of the amphiphiles from/to aggregates of different sizes. We also investigate the dipolar reorientational time correlation function for interfacial water and water molecules in the solvent bulk, as well as the number of hydrogen bonds per molecule in both cases.

Cutaneous T cell lymphoma: translating immunobiology into therapeutic opportunities.

Cutaneous T cell lymphoma (CTCL) has always served as a proving ground where conceptual advances in immunology can be tested and the results translated into clinical practice. From the earliest studies that used sheep red blood cells to identify the malignant cell as a T lymphocyte to molecular demonstration of the clonalilty of the disease, basic science techniques have provided sign posts that allow us to understand the clinical features seen in the patients. We continue to apply this paradigm to develop new insights into the role of the immune system in CTCL with the goal of using this knowledge to enhance the therapeutic options available to the patient. This article will review the studies that have led to our current understanding of the immunobiology of CTCL and the new therapeutic approaches that are being tested in this disease.

Alpha beta and gamma delta T cells can share a late common precursor.

BACKGROUND: The subdivision of T cells into alpha beta and gamma delta subtypes is conserved throughout vertebrate development. The respective alpha beta and gamma delta T-cell receptors (TCRs) are encoded by somatically rearranged genes. There has been broad speculation as to whether an individual thymocyte can become either a gamma delta T cell or an alpha beta T cell as a result of stochastic gene rearrangement processes, or whether the two types of T cell are derived from separate lineages. Many of the experimental findings are apparently conflicting, however, and the issue--a basic one in immunology and development--remains unresolved. RESULTS: To address this issue, we have used the recently developed polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique, which allows us to examine quantitatively the status of TCR gamma and delta genes in postnatal alpha beta T cells and their progenitors. Interestingly, such cells are depleted of productively rearranged delta and gamma genes, which can encode delta and gamma TCR polypeptide chains. However, in mice that can rearrange TCR delta gene segments, but in which the TCR delta gene is non-functional in other respects, no such depletion of productive rearrangements is seen. CONCLUSION: The quantitative data that we have obtained fulfill the predictions of the stochastic hypothesis: that is, a progenitor T cell first attempts to become a gamma delta T cell and, if unsuccessful, then attempts to become an alpha beta T cell. Thus, alpha beta and gamma delta T cells can derive from a common precursor thymocyte. In the simplest case, therefore, lineage-determining factors are the successful rearrangement of both gamma and delta genes before TCR alpha gene rearrangements occur, which lead to deletion of the TCR delta locus and thereby preclude further gamma delta T-cell differentiation. In contrast, successful rearrangement of the TCR beta locus remains compatible with cells becoming either gamma delta or alpha beta T cells.

Specific suppression of lupus-like graft-versus-host disease using extracorporeal photochemical attenuation of effector lymphocytes.

(C57BL/6 x DBA/2)F1 (B6D2F1) mice inoculated with parental DBA/2 (D2) splenocytes develop chronic stimulatory graft-versus-host reaction with many of the clinical manifestations of systemic lupus erythematosus. This investigation tested the ability of 8-methoxypsoralen (8-MOP) and ultraviolet A (UVA) light-treated D2 cells, primed to contain an expanded population of T cells specific for B6D2F1 major histocompatability complex antigens, to treat and/or prevent such systemic lupus erythematosus-like disease. 8-MOP/UVA-treated cells from B6D2F1-primed D2 donors were inoculated into B6D2F1 recipients weekly six to ten times, either before or after initiating graft-versus-host disease with normal D2 cells. A third group of B6D2F1 recipients were vaccinated weekly six times before disease initiation using 8-MOP/UVA-attenuated, B6D2F1-primed D2 cells that had been secondarily stimulated and expanded in vitro in the presence of irradiated B6D2F1 targets and interleukin-2. Control B6D2F1 mice were vaccinated with 8-MOP/UVA-treated D2 cells stimulated in vitro and/or in vivo with (C3H/HeJ x DBA/2)F1 cells. Only mice vaccinated with 8-MOP/UVA-attenuated D2-anti-B6D2F1 cells that were secondarily stimulated and expanded in vitro exhibited differences from controls when measured by the clinical parameters of ascites formation, and mean survival (p < 0.025). These groups also differed significantly in mean antinuclear antibody titer measured 14 weeks after disease initiation (p < 0.05). At 28 weeks, histologic evidence of systemic lupus erythematosus-like kidney disease was found only in the control group. These results indicate that photochemically attenuated D2-anti-B6D2F1 cells primed in vivo and secondarily stimulated and expanded in vitro are capable of vaccinating recipients against progression of graft-versus-host reaction-initiated systemic lupus erythematosus-like disease.

Sensorimotor effects on central space representation: prism adaptation influences haptic and visual representations in normal subjects.

Prism adaptation improves visual and haptic manifestations of left neglect, and can induce a small but reliable simulation of left visual neglect in normal individuals. Here, we present two experiments in which the effects of prism adaptation on the representation of space were explored. In Experiment 1, normal subjects were required to locate the centre of a haptically explored circle, before and after adaptation to leftward displacing prisms. In Experiment 2, a visual circle centring task was used. In both tasks, prism adaptation induced a significant rightward shift of performance. In addition, in both experiments, three classical measures of visuo-manual adaptation were taken: the visual shift, the proprioceptive shift and the total shift. The effects found on the haptic and visual tasks did not correlate with any of these measures. This suggests that the effects of prism adaptation on the circle centring tasks did not depend directly on the sensorimotor consequences of the adaptation. These results imply that prism adaptation can affect noetic levels of space representation in normal subjects, supporting the hypothesis that this low-level sensorimotor intervention can exert a bottom-up structuring influence on higher levels of cognitive integration.

Extracorporeal photochemotherapy in human and murine graft-versus-host disease.

Extracorporeal photochemotherapy (ECP) is an immunotherapy that has found a role in the therapy of cutaneous T cell lymphoma, a disease of mature activated T cells. Graft-versus-host disease (GVHD) is also mediated by activated T cells, and thus often responds to therapies that target T cells. Murine models for both GVHD and ECP can be combined to study the impact of this immunotherapy on GVHD. In this paper we present a patient with GVHD who demonstrated a beneficial therapeutic response to treatment with ECP. The findings of this case are compared with the observations from a murine model for GVHD-ECP. The potential mechanisms of ECP in the treatment of GVHD are discussed. along with the similarities observed with ECP in the treatment of other conditions.

Extracorporeal photochemoimmunotherapy in cutaneous T cell lymphomas.

Extracorporeal photochemotherapy (ECP), or photopheresis, is a widely used treatment for cutaneous T cell lymphoma (CTCL) and other T cell-mediated disorders, having been administered in more than 150 centers worldwide more than 200,000 times. Consistent with the theme of this conference--that is, highlighting the potentially most productive investigative avenues for unraveling the mysteries of CTCL in the next decade--ECP has been futuristic since its inception in the early 1980s. In 1988, the treatment was the first FDA-approved selective immunotherapy for any type of cancer. Yet, the mechanism by which it could suppress a clone of CTCL cells or inactivate multiple autoreactive T cell clones in graft-versus-host disease (GVHD) or allograft rejection remained obscure until quite recently. In fact, the scientific principles necessary to begin to comprehend the basis of ECP's efficacy were not available when the treatment was first introduced in 1982. In the intervening years, necessary detailed knowledge of the structure and function of the clonotypic T cell receptors, of class I major histocompatibility complex (MHC) presentation of tumor antigens, of CTCL tumor-specific antigens, of dendritic antigen presenting cell (DC) biology, and of 8-methoxypsoralen immunopharmacology has been attained. Although much remains to be learned, we now appreciate that ECP simultaneously and efficiently induces both apoptosis of disease-causing T cells and conversion of monocytes to functional DCs. By processing and presenting the unique antigenic determinants of pathogenic T cell clones, the DCs can either initiate a clinically relevant anti-CTCL cytotoxic response or suppress the activity of autoreactive T cell clones. This paper will review clinical trials of ECP in CTCL and evolving scientific understanding of ECP's mechanism in the context of exciting future directions.

The clonotypic T cell receptor is a source of tumor-associated antigens in cutaneous T cell lymphoma.

To develop cancer vaccines for the treatment of cutaneous T cell lymphoma (CTCL), immunogenic peptides were identified by two approaches. First, through the use of "reverse immunology" the peptide sequence of the idiotypic region of the beta chain of the T cell receptor (TCR) was determined and a series of overlapping peptides synthesized and tested for CD8 T cell recognition. In two patients, the idiotypic CDR3 region provided immunogenic epitopes that were recognized in a class I-restricted fashion by autologous CD8 T cell lines. In a second strategy, peptides were isolated directly from class I MHC molecules on the CTCL surface and sequenced. A peptide with partial homology to sequences contained in the conserved variable portion of the clonotypic TCR beta chain was recognized as immunogenic by autologous CD8 T cells. Therefore, both approaches demonstrated that the clonotypic TCR in CTCL is a source of immunogenic tumor epitopes. To confirm that recognition of TCR-derived sequences provides immunoprotection against tumor growth, a murine model of T cell lymphoma was studied. The immunogenicity of a thymoma, which lacks cell surface TCR expression, was enhanced by transfection of the beta chain of the TCR. The studies reviewed in this paper demonstrate that the TCR can serve as one source for immunogenic tumor peptides in T cell lymphoma in vitro and in vivo. Presentation of TCR epitopes on dendritic cells that express high levels of MHC, costimulatory, and adhesion molecules may provide an effective means for immunization against T cell malignancy.

Cutaneous T-cell lymphoma: pathogenesis and treatment.

Cutaneous T-cell lymphoma (CTCL) is a malignancy of a distinctive subset of T-helper cells designated "cutaneous T cells" because of their central role in the normal functioning of the skin immune system. Guided by selective adhesion molecules, activated/memory T cells of the skin immune system normally circulate among the skin, lymph nodes, and peripheral blood. Thus, a better understanding of the skin immune system, which normally functions to provide immunosurveillance against cutaneous pathogens and other insults, has led to a better understanding of the clinical spectrum, pathogenesis, staging, and management of CTCL. This article describes the major subtypes of CTCL and provides an update on the pathogenesis and treatment of this lymphoma.

Balance and aging.

OBJECTIVES/HYPOTHESIS: To provide a basic science and clinical review of normal balance changes with age, and to provide a current review for the evaluation and treatment of elderly patients with balance disorders. As we age, we lose balance function through loss of sensory elements, the ability to integrate information and issue motor commands, and because we lose musculoskeletal function. Diseases common in aging populations lead to further deterioration in balance function in some patients. Treatment of balance dysfunction in aging populations is based on the knowledge of normal aging processes and on an evaluation of the individual's balance loss and remaining balance elements. Prevention and rehabilitation play a major role in treatment; medical and surgical therapy also have a place. STUDY DESIGN AND METHODS: Review of literature, personal research and observations. RESULTS AND CONCLUSIONS: Although older patients may be subject to most of the common balance disorders of younger patients, they have more problems with chronic disequilibrium and falls. Prevention and rehabilitation play an important role in treating these patients.

Predicting fall risks in an elderly population: computer dynamic posturography versus electronystagmography test results.

OBJECTIVES/HYPOTHESIS: Falls are the leading cause of morbidity and mortality for persons aged 65 years and older, with more than 2 million people falling and sustaining serious injury annually. This study compared computer dynamic posturography (CDP) and electronystagmography (ENG) results as predictors of falls. STUDY DESIGN: Retrospective. METHODS: Thirty-three patients over the age of 65 years who presented to a balance disorders and falls prevention clinic were used for this study (22 women and 11 men, with an average age of 78.0 y and a mean fall rate of 3.5 times). All had experienced at least one fall in the year before visiting the clinic and were tested with both CDP and ENG. The CDP results were divided into subcategories (sensory organization testing and limits of stability); ENG results were divided into four categories (ocular motor, rotational chair, positional, and caloric studies). RESULTS: Test findings were classified as normal or abnormal based on age-matched normative data. Of the patients in the study, 27.3% were normal for one type of testing and abnormal for the other. Twenty-six patients (78.8%) had abnormal results on CDP, and 20 individuals (60.6%) showed ENG abnormalities (42.4% for ocular motor, 28.6% for positional, 13.6% for caloric, and 11.2% for rotational chair studies). The limits of stability category was significant in predicting multiple falls. CONCLUSION: For this population, CDP was determined to be a more sensitive test for identifying patients who have fallen, with limits of stability testing the most significant part of the CDP battery; for ENG studies, the best falls indicator was the ocular motor battery.