Novel multispecific heterodimeric antibody format allowing modular assembly of variable domain fragments

Multispecific antibody formats provide a promising platform for the development of novel therapeutic concepts that could facilitate the generation of safer, more effective pharmaceuticals. However, the production and use of such antibody-based multispecifics is often made complicated by: 1) the instability of the antibody fragments of which they consist, 2) undesired inter-subunit associations, and 3) the need to include recombinant heterodimerization domains that confer distribution-impairing bulk or enhance immunogenicity. In this paper, we describe a broadly-applicable method for the stabilization of human or humanized antibody Fv fragments that entails replacing framework region IV of a V_κ1/V_H3-consensus Fv framework with the corresponding germ-line sequence of a λ-type V_L chain. We then used this stable Fv framework to generate a novel heterodimeric multispecific antibody format that assembles by cognate V_L/V_H associations between 2 split variable domains in the core of the complex. This format, termed multispecific antibody-based therapeutics by cognate heterodimerization (MATCH), can be applied to produce homogeneous and highly stable antibody-derived molecules that simultaneously bind 4 distinct antigens. The heterodimeric design of the MATCH format allows efficient in-format screening of binding domain combinations that result in maximal cooperative activity.     

Laser-assisted targeted gene delivery to degenerated retina improves retinal function

Delivery of therapeutic genes into retina is proving to reverse degeneration and restore vision, however, viral vector-based gene delivery is prone to immunorejection, inflammatory/immune-response and nontargeted. Here, we report nonviral gene delivery and expression of opsin encoding genes in mouse retina in-vitro and in-vivo by use of pulsed femtosecond laser microbeam. In-vitro patch-clamp recording of the opsin-sensitized retinal cells and visually evoked in-vivo electrical recording from laser-transfected eye of mouse with degenerated retina showed functional response. The ultrafast laser-based naked gene delivery showed minimal damage and reliable expression of therapeutic opsin in cell membrane of the selected cells and in targeted retinal region. Laser-based “naked DNA gene therapy” in a spatially targeted manner will pave the way for treatment of inherited retinal diseases.     

Potentiation of the efficacy of murine L1210 leukemia vaccine by a novel immunostimulator 7-thia-8-oxoguanosine: Increased survival after immunization with vaccine plus 7-thia-8-oxoguanosine

Summary We have investigated the ability of a novel immunopotentiator, 7-thia-8-oxoguanosine (7T8OG) to increase the efficacy of a weakly immunogenic murine L1210 leukemia vaccine. The vaccine was prepared by irradiating L1210 leukemia cells in a cesium source with a total of 6000-R dose. DBA/2 mice were treated with 150 mg/kg 7T8OG and/or with vaccine consisting of 10⁷ irradiated cells. In combination therapy, mice first received the vaccine and then were injected with 75 mg/kg 7T8OG 2 h and 4 h after vaccination. One week after the last treatment all mice were inoculated with 10⁴ live leukemia cells intraperitoneally. Control, untreated mice (n = 66) injected with 10⁴ live leukemia cells had a mean survival time ± standard error of 10.5±0.2 days. Treating mice (n = 66) with one, two or three doses of 7T8OG administered i.p. 1 week apart did not increase survival (mean survival time = 10.7 days). Mice immunized with one, two or three doses of vaccine had 14.5±1.1, 45.4±6.2 and 68.3±10.6 days mean survival, respectively. 7T8OG-stimulated vaccination increased the survival dramatically. The best survival was noted when the mice were treated with 2× (vaccine + 7T8OG). Immunization of mice (n = 30) with this treatment regimen increased the mean survival to 156±10.0 days. Over 90% of mice that were treated this way had a cumulative survival time greater than 160 days. In contrast, only 12% of the mice immunized twice with the leukemia vaccine alone survived over 160 days. These results suggest a rationale for the use of this immuno-potentiator with various vaccines for a more effective immunization.     

AChemS: the beginning. Association for Chemoreception Sciences

This paper unfolds the events, the people and the times that led up to the founding of AChemS and fashioned its character during its early formative years. It describes the path over which AChemS came, going from the original assertions and denials for the need of such an organization to its later inception and nascent development. This narration highlights such topics as the debate over the need for AChemS, the role of National Science Foundation in the founding of AChemS, the derivation of the Association's name, the choice of Sarasota and the Hyatt House as the meeting site, the generation of the programs for the early annual meetings, the adoption of the bylaws, the process of incorporation and tax deferment, and the birth of the Givaudan Lectureship. Most emphatically highlighted, however, is the enthusiasm, commitment and hard work that the members of the chemosensory research community displayed in bringing AChemS to fruition.      

Glycoproteins in Rathke's gland secretions of loggerhead (Caretta caretta) and Kemp's ridley (Lepidochelys kempi) sea turtles

1. The Rathke's gland secretions of loggerhead (Caretta caretta) and Kemp's ridley (Lepidochelys kempi) sea turtles contain 20 and 10 mg of protein/ml, respectively. The proteins of each species were separated by gel filtration into two major fractions, one (35%) in the excluded volume, and one (50%) with a molecular mass of approximately 55 kDA. 2. The 55 kDa fraction from each species' secretions exhibits a single band on SDS-PAGE (Mr approximately equal to 55,000) and a single amino-terminal sequence. 3. The amino acid compositions of the two 55 kDa proteins are similar, and the first 15 residues of their amino terminus are identical. Both proteins contain glucosamine. 4. Analyses of the amino acid and amino sugar composition of the high molecular weight fractions from the two turtle species also indicate similarities; there are distinct differences between them and their 55 kDa proteins.     

Bioconversion of poultry manure and vegetable waste mixes by Hermetia illucens (Diptera: Stratiomyidae)

Development time, survival and final nutrient content of Hermetia illucens L. larvae depends on the substrates in which they develop. Mixing different waste types together can increase the performance and survival of the larvae, as well as their waste reduction. The main objective of this study was to evaluate the effect of different ratios of mixed fruit and vegetable waste with poultry manure on larval development time, size, biomass production, survival, bioconversion and waste reduction. Freshly hatched neonates (90 mg; approx. 6000 individuals) were placed on 12 kg of a mixture of fruit and vegetable waste and fresh, unprocessed poultry manure and held at 28 ± 0.5°C. Inclusion of fruit and vegetable waste varied from 0% to 100% in 10% increments. Initial temperature of the substrate was also measured. The individual mass of larvae increased significantly as more fruit and vegetable waste was included, from less than 81.3 ± 6.6 mg on poultry manure only to an average size of 211.6 ± 6.0 mg at 100% fruit and vegetable waste. After approximately 60% inclusion of fruit and vegetable waste the performance and survival of the larvae increased significantly while development time was reduced. A combination of high fruit and vegetable waste and low initial temperatures resulted in lower development time overall. The mixing of wastes can be applied in industry to further the goals of waste reduction and biomass production while incorporating low-quality wastes like poultry manure.     

Precipitation contributes to plant height,but not reproductive effort,for western prairie fringed orchid (Platanthera praeclara Sheviak & Bowles): Evidence from herbarium records

The western fringed prairie orchid (WFPO) is a rare plant found in mesic to wet tallgrass prairies in the Great Plains and Midwest regions of the United States. The size of WFPO populations varies considerably from year to year, and studies have suggested that population size is dependent on precipitation during critical periods in the plant's annual development. We hypothesized that plant height and reproductive effort would also be controlled by precipitation, either during these periods or over a broader period. We acquired available images of WFPO from 21 herbaria, and of these 141 individual plants had information adequate for analysis, although some population/year combinations were represented multiple times. For each specimen, we measured plant height (cm) and reproductive effort (as measured by total flower and bud count). We used bootstrapped linear regression, randomly selecting one individual from each population/year combination, to compare precipitation models, both during critical periods and the various summaries. We found that precipitation during the phenologically critical periods was a poor predictor of plant height and reproductive effort. Of the broader precipitation variables, accumulated precipitation from January 1 to collection date best described plant height. We also used correlations to detect a relationship among the variables WFPO height, reproductive effort, precipitation, latitude, and year of collection. Year of specimen collection was negatively correlated with WFPO plant height and accumulated precipitation, suggesting that both have declined in more recent years. Negative correlations with latitude also suggest height and precipitation decrease in the northern part of WFPO's range. Reproductive effort was not related to tested precipitation variables; however, it was weakly correlated with plant height. Although the results are limited, this study leverages available data and makes inferences on WFPO biology over broad ranges of time (1894–2012) and latitude (37.5°–49.9°).