Polarization state generation and measurement with a single metasurface.

The constituent elements of metasurfaces may be designed with explicit polarization dependence, making metasurfaces a fascinating platform for new polarization optics. In this work we show that a metasurface grating can be designed to produce arbitrarily specified polarization states on a set of defined diffraction orders given that the polarization of the incident beam is known. We also demonstrate that, when used in a reverse configuration, the same grating may be used as a parallel snapshot polarimeter, requiring a minimum of bulk polarization optics. We demonstrate its use in measuring partially polarized light, and show that it performs favorably in comparison to a commercial polarimeter. This work is of consequence in any application requiring lightweight, compact, and low-cost polarization optics, polarimetry, or polarization imaging.

Lateral chirality-sorting optical forces.

The transverse component of the spin angular momentum of evanescent waves gives rise to lateral optical forces on chiral particles, which have the unusual property of acting in a direction in which there is neither a field gradient nor wave propagation. Because their direction and strength depends on the chiral polarizability of the particle, they act as chirality-sorting and may offer a mechanism for passive chirality spectroscopy. The absolute strength of the forces also substantially exceeds that of other recently predicted sideways optical forces.

Metasurface Polarization Optics: Independent Phase Control of Arbitrary Orthogonal States of Polarization.

We present a method allowing for the imposition of two independent and arbitrary phase profiles on any pair of orthogonal states of polarization-linear, circular, or elliptical-relying only on simple, linearly birefringent wave plate elements arranged into metasurfaces. This stands in contrast to previous designs which could only address orthogonal linear, and to a limited extent, circular polarizations. Using this approach, we demonstrate chiral holograms characterized by fully independent far fields for each circular polarization and elliptical polarization beam splitters, both in the visible. This approach significantly expands the scope of metasurface polarization optics.

Community-based livestock breeding programmes: essentials and examples.

Breeding programmes described as community-based (CBBP) typically relate to low-input systems with farmers having a common interest to improve and share their genetic resources. CBBPs are more frequent with keepers of small ruminants, in particular smallholders of local breeds, than with cattle, pigs or chickens with which farmers may have easier access to alternative programmes. Constraints that limit the adoption of conventional breeding technologies in low-input systems cover a range of organizational and technical aspects. The analysis of 8 CBBPs located in countries of Latin-America, Africa and Asia highlights the importance of bottom-up approaches and involvement of local institutions in the planning and implementation stages. The analysis also reveals a high dependence of these programmes on organizational, technical and financial support. Completely self-sustained CBBPs seem to be difficult to realize. There is a need to implement and document formal socio-economic evaluations of CBBPs to provide governments and other development agencies with the information necessary for creating sustainable CBBPs at larger scales.

Polarization-selective coupling to long-range surface plasmon polariton waveguides.

We demonstrate polarization-selective coupling from an optical fiber to long-range surface plasmon polariton waveguide modes using plasmonic antenna arrays. The arrays allow the sorting of two distinct (not necessarily orthogonal) polarizations to counter-propagating waveguide modes. The polarization-selective behavior of the devices is described by a compact formalism based on Stokes vectors that offers a clear graphical representation of the response. We experimentally observe polarization-controlled switching and unidirectional coupling with extinction ratios greater than 30 dB and coupling efficiencies comparable to those of a conventional grating coupler.

Converting multi-trait breeding objectives into operative selection indexes to ensure genetic gains in low-input sheep and goat breeding programmes.

Small ruminant breeding programmes in low-input production systems are best organised at the community level. Participant farmers have to agree on goal traits and their relative importance. When BLUP breeding values of goal traits are not available in time, appropriate selection indexes can be used to aid visual selection. Taking Ethiopian Abergelle goat and Bonga sheep community-based breeding programmes (CBBPs) as an example, breeding objective functions were defined and selection indexes were constructed and evaluated. Breeding goals for Abergelle goats included early sale weight, survival and milk production. Breeding goals for Bonga included the number of offspring born, sale weight and survival. Economic weights of objective traits can be used in several ways depending on measured traits and the reliability of their genetic parameters. Selection indexes included combinations of objective traits measured on candidates and their dams and situations when Abergelle communities prefer to restrict genetic changes in number of offspring born or adult weight and when Bonga communities prefer to restrict changes in adult weight. Genetic and economic gains were evaluated as well as sensitivity to feed cost assumptions and to repeated dam records. After independent culling on preponderant traits such as coat colour and horn/tail type, sires in Abergelle goat community breeding programmes should be selected on indexes including at least own early live weight and their dams average milk production records. Sires for Bonga sheep programmes should be selected on own early live weight and desirably also on their dam's number of offspring born. Sensitivity to feed cost assumptions was negligible but repeated measurements of dam records improved index accuracies considerably. Restricting genetic changes in number of offspring born or adult weight is not recommended.

Community-based sheep breeding programs generated substantial genetic gains and socioeconomic benefits.

Community-based breeding programs (CBBPs) for small ruminants have been suggested as alternatives to centralised, government-controlled breeding schemes which have been implemented in many developing countries. An innovative methodological framework on how to design, implement and sustain CBBPs was tested in three sites in Ethiopia: Bonga, Horro and Menz. In these CBBPs, the main selection trait identified through participatory approaches was 6-month weight in all three sites. In Horro and Bonga, where resources such as feed and water permitted larger litter sizes, twinning rate was included. Ten-year (2009 to 2018) performance data from the breeding programs were analysed using Average Information Restricted Maximum Likelihood method (AI-REML). Additionally, the socioeconomic impact of CBBPs was assessed. Results indicated that 6-month weight increased over the years in all breeds. In Bonga, the average increase was 0.21 ± 0.018 kg/year, followed by 0.18 ± 0.007 and 0.11 ± 0.003 kg/year in Horro and Menz, respectively. This was quite substantial in an on-farm situation. The birth weight of lambs did not improve over the years in Bonga and Horro sheep but significant increases occurred in Menz. Considering that there was no direct selection on birth weight in the community flock, the increased weights observed in Menz could be due to correlated responses, but this was not the case in Bonga and Horro. The genetic trend for prolificacy over the years in both Bonga and Horro flocks was positive and significant (P < 0.01). This increase in litter size, combined with the increased 6-month body weight, increased income by 20% and farm-level meat consumption from slaughter of one sheep per year to three. The results show that CBBPs are technically feasible, result in measurable genetic gains in performance traits and impact the livelihoods of farmers.

High-efficiency chiral meta-lens.

We present here a compact metasurface lens element that enables simultaneous and spatially separated imaging of light of opposite circular polarization states. The design overcomes a limitation of previous chiral lenses reliant on the traditional geometric phase approach by allowing for independent focusing of both circular polarizations without a 50% efficiency trade-off. We demonstrate circular polarization-dependent imaging at visible wavelengths with polarization contrast greater than 20dB and efficiencies as high as 70%.

Treatment of experimental encephalomyelitis with a novel chimeric fusion protein of myelin basic protein and proteolipid protein.

It has been shown that peripheral T cell tolerance can be induced by systemic antigen administration. We have been interested in using this phenomenon to develop antigen-specific immunotherapies for T cell-mediated autoimmune diseases. In patients with the demyelinating disease multiple sclerosis (MS), multiple potentially autoantigenic epitopes have been identified on the two major proteins of the myelin sheath, myelin basic protein (MBP) and proteolipid protein (PLP). To generate a tolerogenic protein for the therapy of patients with MS, we have produced a protein fusion between the 21.5-kD isoform of MBP (MBP21.5) and a genetically engineered form of PLP (deltaPLP4). In this report, we describe the effects of treatment with this agent (MP4) on clinical disease in a murine model of demyelinating disease, experimental autoimmune encephalomyelitis (EAE). Treatment of SJL/J mice with MP4 after induction of EAE either by active immunization or by adoptive transfer of activated T cells completely prevented subsequent clinical paralysis. Importantly, the administration of MP4 completely suppressed the development of EAE initiated by the cotransfer of both MBP- and PLP-activated T cells. Prevention of clinical disease after the intravenous injection of MP4 was paralleled by the formation of long-lived functional peptide-MHC complexes in vivo, as well as by a significant reduction in both MBP- and PLP-specific T cell proliferative responses. Mice treated with MP4 were resistant to disease when rechallenged with an encephalitogenic PLP peptide emulsified in CFA, indicating that MP4 administration had a prolonged effect in vivo. Administration of MP4 was also found to markedly ameliorate the course of established clinical disease. Finally, MP4 therapy was equally efficacious in mice defective in Fas expression. These results support the conclusion that MP4 protein is highly effective in suppressing disease caused by multiple neuroantigen epitopes in experimentally induced demyelinating disease.

Rad23 is required for transcription-coupled repair and efficient overrall repair in Saccharomyces cerevisiae.

The repair of UV-induced photoproducts (cyclobutane pyrimidine dimers) in a well-characterized minichromosome, genomic DNA, and a transcribed genomic gene (RPB2) of a rad23delta mutant of Saccharomyces care was examined. Isogenic wild-type cells show a strong bias for the repair of the transcribed strands in both the plasmid and genomic genes and efficient overall repair of both DNAs (>80% of the dimers were removed in 6 h). However, the rad23delta mutant shows (i) no strand bias for repair in these genes and decreased repair of both strands, (ii) partial repair of genomic DNA (approximately 45% in 6 h), and (iii) very poor repair of the plasmid overall approximately 15% in 6 h). These features, coupled with the decreased UV survival of rad23delta cells, indicate that Rad23 is required for both transcription-coupled repair and efficient overall repair in S. cerevisiae.

Arbitrary spin-to-orbital angular momentum conversion of light.

Optical elements that convert the spin angular momentum (SAM) of light into vortex beams have found applications in classical and quantum optics. These elements-SAM-to-orbital angular momentum (OAM) converters-are based on the geometric phase and only permit the conversion of left- and right-circular polarizations (spin states) into states with opposite OAM. We present a method for converting arbitrary SAM states into total angular momentum states characterized by a superposition of independent OAM. We designed a metasurface that converts left- and right-circular polarizations into states with independent values of OAM and designed another device that performs this operation for elliptically polarized states. These results illustrate a general material-mediated connection between SAM and OAM of light and may find applications in producing complex structured light and in optical communication.

Adhesion of acidic lipid vesicles by 21.5 kDa (recombinant) and 18.5 kDa isoforms of myelin basic protein.

Myelin basic protein (MBP) is thought to be responsible for adhesion of the intracellular surfaces of compact myelin to give the major dense line. The 17 and 21.5 kDa isoforms containing exon II have been reported by others to localize to the cytoplasm and nucleus of murine oligodendrocytes and HeLa cells while the 14 and 18.5 kDa isoforms lacking exon II are confined to the plasma membrane. However, we show that the exon II(-) 18.5 kDa form and a recombinant exon II(+) 21.5 kDa isoform both caused similar aggregation of acidic lipid vesicles, indicating that they should have similar abilities to bind to the intracellular lipid surface of the plasma membrane and to cause adhesion of those surfaces to each other. The circular dichroism spectra of the two isoforms indicated that both had a similar secondary structure. Thus, both isoforms should be able to bind to and cause adhesion of the cytosolic surfaces of compact myelin. The fact that they do not could be due to differences in post-translational modification in vivo, trafficking through the cell and/or subcellular location of synthesis, but it is not due to differences in their lipid binding.

Purification of immunologically active recombinant 21.5 kDa isoform of human myelin basic protein.

We have designed and expressed in bacteria a recombinant fetal form of human myelin basic protein (21.5 kDa isoform; rhMBP21.5), a candidate autoantigen in multiple sclerosis. An exon 2 insertion, carboxy-terminal histidine tag and preferred bacterial codons differentiate the MBP21.5 gene from that encoding the adult, brain-derived form of human MBP (18.5 kDa isoform; hMBP18.5). MBPs were expressed at high levels in E. coli and extracted from whole cells by simultaneous acid solubilization and mechanical disruption. A nearly two-fold increase in recombinant protein was detected in strains harboring MBP genes with bacterial preferred codons compared to genes containing human codons. The recombinant molecules were purified in two steps, first by reversed-phase chromatographic separation and then by metal affinity chromatography. Dimeric forms of recombinant MBP21.5 were detected under physiological conditions, however, substitution of a serine for the single cysteine at amino acid residue 81 resulted in only monomer formation. All forms of recombinant MBPs induced proliferative responses of human T lymphocytes specific for epitopes in MBP18.5 kDa. In contrast, human T cell lines that recognize an exon 2-encoded epitope of MBP responded to the 21.5 kDa isoform of MBP, but not the 18.5 kDa isoform.

Humanized porcine VCAM-specific monoclonal antibodies with chimeric IgG2/G4 constant regions block human leukocyte binding to porcine endothelial cells.

Discordant xenografts surviving the initial hyperacute rejection phase may be subject to cellular rejection processes mediated by infiltrating leukocytes including T cells, NK cells and monocytes. The stable adhesion of these cell types to endothelial cells is due to the molecular interaction of the integrins VLA-4 and LFA-1 with their ligands vascular cell adhesion molecule (VCAM) and ICAM-1 present on the endothelial cells. Human VLA-4 binds to porcine VCAM, and blocking mAbs specific for porcine VCAM have been developed. We have localized the epitope of the anti-porcine VCAM blocking mAbs 2A2 and 3F4 to domains 1 and 2, respectively. Humanized antibodies (IgG4 isotype) were constructed from these anti-porcine VCAM antibodies and demonstrated to inhibit adhesion of Ramos, Jurkat and YT cells, as well as purified resting and activated human T cells, to porcine aortic endothelial cells (PAEC). These cell types express both LFA-1 as well as VLA-4, suggesting blockade of human VLA-4 interaction with porcine VCAM may alone be sufficient to significantly impair adhesion of human leukocytes to porcine endothelial cells. The chimeric anti-porcine VCAM (pVCAM) HuG4 antibodies promoted increased adhesion of Fc receptor (FcR) positive cells such as U937 monocytic cells to PAEC. In contrast, chimeric anti-porcine VCAM antibodies created using the CH1 and hinge region from human IgG2 and the CH2 and CH3 regions from human IgG4 (HuG2/G4 antibodies) inhibited binding of FcR positive cells to PAEC. These chimeric anti-pVCAM antibodies should allow delineation of the in vivo role of VLA-4/VCAM interaction in porcine-to-primate xenotransplants. Further, the design of the HuG2/G4 antibodies should render them efficacious in multiple settings requiring elimination of FcR binding.

In vitro and in vivo inhibition of complement activity by a single-chain Fv fragment recognizing human C5.

Complement activation has been implicated in the pathogenesis of several human diseases. Recently, a monoclonal antibody, (N19-8) that recognizes the human complement protein C5 has been shown to effectively block the cleavage of C5 into C5a and C5b, thereby blocking terminal complement activation. In this study, a recombinant N19-8 scFv antibody fragment was constructed from the N19-8 variable regions, and produced in both mammalian and bacterial cells. The N19-8 scFv bound human C5 and was as potent as the N19-8 monoclonal antibody at inhibiting human C5b-9-mediated hemolysis of chicken erythrocytes. In contrast, the N19-8 scFv only partially retained the ability of the N19-8 monoclonal antibody to inhibit C5a generation. To investigate the ability of the N19-8 scFv to inhibit complement-mediated tissue damage, complement-dependent myocardial injury was induced in isolated mouse hearts by perfusion with Krebs-Henseleit buffer containing 6% human plasma. The perfused hearts sustained extensive deposition of human C3 and C5b-9, resulting in increased coronary artery perfusion pressure, end-diastolic pressure, and a decrease in heart rate until the hearts ceased beating approximately 10 min after addition of plasma. Hearts treated with human plasma supplemented with either the N19-8 monoclonal antibody or the N19-8 scFv did not show any detectable changes in cardiac performance for at least 1 hr following the addition of plasma. Hearts treated with human plasma alone showed extensive deposition of C3 and C5b-9, while hearts treated with human plasma containing N19-8 scFv showed extensive deposition of C3, but no detectable deposition of C5b-9. Administration of a 100 mg bolus dose of N19-8 scFv to rhesus monkeys inhibited the serum hemolytic activity by at least 50% for up to 2 hr. Pharmacokinetic analysis of N19-8 scFv serum levels suggested a two-compartment model with a T1/2 alpha of 27 min. Together, these data suggest the recombinant N19-8 scFv is a potent inhibitor of the terminal complement cascade and may have potential in vivo applications where short duration inhibition of terminal complement activity is desirable.

Porcine vascular cell adhesion molecule (VCAM) mediates endothelial cell adhesion to human T cells. Development of blocking antibodies specific for porcine VCAM.

Vascular cell adhesion molecule (VCAM) is expressed on activated endothelial cells and binds to the alpha 4 beta 1 integrin receptor, very late antigen-4 (VLA-4), expressed on human lymphoid cells. Anti-VCAM mAbs have been shown to prolong allograft survival. To explore the role of porcine VCAM (pVCAM) in xenotransplantation, a recombinant secreted form of pVCAM (spVCAM) was expressed in 293-EBNA cells and purified by metal affinity chromatography. A human lymphoid cell line bound to spVCAM in a VLA-4-dependent manner. Using spVCAM as an immunogen, we developed three anti-pVCAM mAbs that reacted with cell surface pVCAM on porcine aortic endothelial cells (PAEC) but not to human VCAM on human umbilical vein endothelial cells. Pairwise interaction analysis indicated that these mAbs recognized distinct epitopes on pVCAM. Two anti-pVCAM mAbs, 2A2 and 3F4, inhibited the binding of Ramos cells to spVCAM, while the third, 5D11, did not. Similarly, mAbs 2A2 and 3F4 inhibited binding of Ramos cells or human peripheral blood T cells to activated PAEC. The extent of inhibition with mAbs 2A2 and 3F4 was comparable to the inhibition obtained with a blocking mAb to human VLA-4. These anti-pVCAM mAbs will provide a means to specifically block pVCAM in a xenograft setting and allow the determination of the role of pVCAM in a primary xenogeneic immune response.

Immune tolerance mediated by recombinant proteolipid protein prevents experimental autoimmune encephalomyelitis.

Proteolipid protein (PLP), a transmembrane protein expressed only in the central nervous system (CNS), is a candidate target autoantigen for autoimmune-mediated demyelination. We have evaluated the effect of a recombinant form of the PLP protein, delta PLP4, in a murine model of experimental autoimmune encephalomyelitis (EAE). PLP-specific T-cell responses were observed following immunization of SJL/J, PL/J and SWR mice with delta PLP4, demonstrating processing of the protein to several distinct antigenic epitopes. Clinical EAE associated with inflammation and demyelination in the CNS also developed after sensitization of mice with delta PLP4 in adjuvant. Conversely, tolerance to delta PLP4 in adult mice and prevention of PLP peptide 139-151-induced EAE was induced by intravenous injection of soluble delta PLP4. The prevention of disease onset was paralleled by a significant reduction in demyelination and CNS inflammatory cell infiltration and diminished PLP139-151-specific T-cell proliferative responses. These results are consistent with the establishment of peripheral T-cell tolerance and reinforce the notion that recombinant myelin antigens and intravenous tolerance induction may prove useful in the modulation of the human demyelinating disease, multiple sclerosis (MS).

Identification of Bacillus subtilis adaptive response genes by subtractive differential hybridization.

Subtractive differential hybridization was used to identify genes in Bacillus subtilis that are induced by nutrient limitation. Several transcription units were identified. They exhibited increased transcription when cells were deprived of certain nutrients, such as glucose, ammonium, or phosphate, or when cells were treated with decoyinine. The genes have been designated dci (for decoyinine-inducible) and gsi (for glucose-starvation-inducible). Using lacZ transcriptional fusions, the dependence of dci and gsi expression on gene products of the sensor and activator classes of bacterial two-component regulatory systems was examined. Transcription of dciA was impaired by a mutation in spoOA, while expression of gsiA was dependent on the early competence genes comP and comA. The implications of these findings are discussed, and a provisional scheme for information flow during the transition phase from growth to sporulation is proposed.

Amelioration of autoimmune reactions by antigen-induced apoptosis of T cells.

We have shown that T cells vigorously cycling in response to growth lymphokines are driven into apoptosis by potent TCR restimulation. This process, termed propriocidal regulation, appears to be a normal feedback inhibitory mechanism to prevent excessive T cell proliferation and lymphokine production. Exposure of T cells to repeated high dose antigen treatments creates the conditions just described by activating T cells, and stimulating the production of growth lymphokines and their receptors. High growth lymphokine levels induced by the large amount of antigen present, stimulate vigorous cycling. The continued presence of high antigen levels subjects the cycling T cells to strong TCR restimulation as they enter the vulnerable S phase, inducing apoptosis in T cells responsive to the administered antigen. Thus, simple, repetitive, intravenous administration of high dose antigen may be used to delete potentially destructive clones of T cells, resulting in a state of peripheral tolerance. This has obvious therapeutic potential in disorders where the elimination of pathogenic T cell clones could be beneficial. We have described in EAE, an animal model for MS, that high dose MBP therapy is effective in preventing CNS pathology and the onset of disease as well as reducing the severity of the clinical symptoms of established EAE. We are currently involved in expanding this approach to other animal models of autoimmunity and graft rejection, as well as refining the immunotherapy in the EAE model with the objective of developing a clinical therapy for human demyelinating disease.

Forage characteristics, steer performance, and water quality from bermudagrass pastures fertilized with two levels of nitrogen from swine lagoon effluent.

Four .8-ha pastures of bermudagrass (Cynodon dactylon [L.] Pers.) were fertilized with either 456 or 873 kg/ha of nitrogen (N) from swine lagoon effluent (two replicates per treatment) and grazed by steers over two summers. Within each pasture, steers received forage only, an energy source (corn), a mixture of corn and soybean meal, or a mixture of corn and blood meal via electronic Calan feeders. All supplements were offered at a level of 1.36 kg/d, and the soybean meal and blood meal supplements provided similar among quantities of protein. Weight gains were similar among supplemented steers, but supplemented steers gained faster (P < .05) than controls. Nitrogen fertilization level had no effect on steer gains, steer grazing days per hectare, or in vitro dry matter disappearance, NDF, and ADF of clipped forage samples. Plant protein and nitrate ion concentrations were greater (P < .06) in clipped forage samples receiving the higher N application rate. Nitrate ion concentrations were greater in available forage samples from the pastures with the high N application rate. Mean total N and nitrate N concentrations were similar in water samples obtained from monitoring wells for the two N treatments over the 2 yr and there were no year x N interactions. Chloride concentrations were greater (P < .05) and pH and specific conductance were less in water samples collected from the 873 kg than from the 456 kg/ha N treatment. Long-term studies are needed to examine the possible cumulative effects of applying various levels of swine waste to the same land area.