The CEA/CD3-bispecific antibody MEDI-565 (MT111) binds a nonlinear epitope in the full-length but not a short splice variant of CEA

MEDI-565 (also known as MT111) is a bispecific T-cell engager (BiTE?) antibody in development for the treatment of patients with cancers expressing carcinoembryonic antigen (CEA). MEDI-565 binds CEA on cancer cells and CD3 on T cells to induce T-cell mediated killing of cancer cells. To understand the molecular basis of human CEA recognition by MEDI-565 and how polymorphisms and spliced forms of CEA may affect MEDI-565 activity, we mapped the epitope of MEDI-565 on CEA using mutagenesis and homology modeling approaches. We found that MEDI-565 recognized a conformational epitope in the A2 domain comprised of amino acids 326-349 and 388-410, with critical residues F(326), T(328), N(333), V(388), G(389), P(390), E(392), I(408), and N(410). Two non-synonymous single-nucleotide polymorphisms (SNPs) (rs10407503, rs7249230) were identified in the epitope region, but they are found at low homozygosity rates. Searching the National Center for Biotechnology Information GenBank? database, we further identified a single, previously uncharacterized mRNA splice variant of CEA that lacks a portion of the N-terminal domain, the A1 and B1 domains, and a large portion of the A2 domain. Real-time quantitative polymerase chain reaction analysis of multiple cancers showed widespread expression of full-length CEA in these tumors, with less frequent but concordant expression of the CEA splice variant. Because the epitope was largely absent from the CEA splice variant, MEDI-565 did not bind or mediate T-cell killing of cells solely expressing this form of CEA. In addition, the splice variant did not interfere with MEDI-565 binding or activity when co-expressed with full-length CEA. Thus MEDI-565 may broadly target CEA-positive tumors without regard for expression of the short splice variant of CEA. Together our data suggest that MEDI-565 activity will neither be impacted by SNPs nor by a splice variant of CEA.     

Use of the 'Food Vacuole Content' Method to Estimate Grazing by the Mixotrophic Dinoflagellate Gyrodinium Galatheanum on Cryptophytes

We measured in situ grazing rates of the mixotrophic dinoflagellateGyrodinium galatheanum (Braarud) Taylor 1995 on populationsof phycoerythrin-containing cryptophytes in Chesapeake Bay.Rates were estimated from instantaneous food vacuole contents,in situ temperatures, cryptophyte abundances and experimentallydetermined digestion rates. Laboratory digestion experimentsshowed that specific digestion rate constants increased sigmoidallywith temperature, but were unrelated to the initial food vacuolecontent when it was <0.46 cryptophytes dinoflagellate^–1.These results allowed us to establish an empirical model toestimate in situ ingestion of cryptophyte prey by G. galatheanum.The estimated rates ranged from 0 to 0.26 cryptophytes dinoflagellate^–1day^–1, corresponding to daily ingestion of 0–12.29pg carbon, 0–2.48 pg nitrogen and 0–0.34 pg phosphorusdinoflagellate^–1. Estimated daily consumption of cryptophytebiomass by G. galatheanum was equivalent to 0–12% of bodycarbon, 0–13% of body nitrogen and 0–21% of bodyphosphorus. Estimated in situ clearance rates for cryptophytesranged from 0 to 0.27 µl dinoflagellate^–1 day^–1,representing daily removal of 0–4% of the cryptophytestanding stock. Although G. galatheanum may increase its growthrate through phagotrophy, it appears to have little grazingimpact on cryptophyte prey populations.     

Revision of the family Duboscquellidae with description of Euduboscquella crenulata n. gen., n. sp. (Dinoflagellata, Syndinea), an intracellular parasite of the ciliate Favella panamensis Kofoid & Campbell

Recent recognition that tintinnids are infected by dinophycean as well as syndinean parasites prompts taxonomic revision of dinoflagellate species that parasitize these ciliates. Long overlooked features of the type species Duboscquella tintinnicola are used to emend the genus and family Duboscquellidae, resulting in both taxa being moved from the Syndinea to the Dinophyceae. Syndinean species previously classified as Duboscquella are relocated to Euduboscquella n. gen., with Euduboscquella crenulata n. sp. as the type. As an endoparasitic species, E. crenulata shares with its congeners processes associated with intracellular development and sporogenesis, but differs from closely related species in nuclear and cortical morphology of the trophont, including a distinctively grooved shield (= episome) that imparts a crenulated appearance in optical section. In addition, E. crenulata produces three morphologically distinct spore types, two of which undergo syngamy to form a uninucleate zygote. The zygote undergoes successive division to produce four daughter cells of unequal size, but that resemble the nonmating spore type.     

Telotroch formation, survival, and attachment in the epibiotic peritrich Zoothamnium intermedium (Ciliophora, Oligohymenophorea)

Abstract. Aspects of the life cycle of the peritrich ciliate Zoothamnium intermedium , an epibiont on calanoid copepods in the Chesapeake Bay, were investigated using host and epibiont cultures. Experiments were designed to characterize the formation, survival, and attachment of free-swimming stages (telotrochs) and to assess whether telotrochs preferentially attach to primary ( Acartia tonsa and Eurytemora affinis ) or alternate hosts from the zooplankton community (the rotifer Brachionus plicatilis , barnacle nauplii, polychaete larvae, and a harpacticoid copepod). The results showed that telotroch formation started 2 h after the death of the host, with >90% of the zooids leaving the host carapace within 7 h. Formation of telotrochs was triggered only by the death of the host, failing to occur when the host was injured or unable to swim. Telotrochs failed to attach to non-living substrates and survived for only 14 h in the absence of host organisms, suggesting that members of Z. intermedium are obligate epibionts. Attachment success decreased with telotroch age, indicating that colonization success in nature may strongly depend on the ability to find a suitable host in a short period of time. Individuals exhibited no preferences in colonizing juvenile or adult stages of A. tonsa or E. affinis . While telotrochs were able to colonize barnacle nauplii and the harpacticoid copepod in the absence of individuals of A. tonsa or E. affinis , they did not attach to the rotifers or polychaete larvae. Telotrochs preferentially colonized individuals of A. tonsa when in the presence of other non-calanoid host species.     

Differential Resistance among Host and Non-host Species Underlies the Variable Success of the Hemi-parasitic Plant Rhinanthus minor

Annals of Botany      

White matter tract-oriented deformation predicts traumatic axonal brain injury and reveals rotational direction-specific vulnerabilities

A systematic correlation between finite element models (FEMs) and histopathology is needed to define deformation thresholds associated with traumatic brain injury (TBI). In this study, a FEM of a transected piglet brain was used to reverse engineer the range of optimal shear moduli for infant (5 days old, 553–658 Pa) and 4-week-old toddler piglet brain (692–811 Pa) from comparisons with measured in situ tissue strains. The more mature brain modulus was found to have significant strain and strain rate dependencies not observed with the infant brain. Age-appropriate FEMs were then used to simulate experimental TBI in infant (\(n=36\)) and preadolescent (\(n=17\)) piglets undergoing a range of rotational head loads. The experimental animals were evaluated for the presence of clinically significant traumatic axonal injury (TAI), which was then correlated with FEM-calculated measures of overall and white matter tract-oriented tissue deformations, and used to identify the metric with the highest sensitivity and specificity for detecting TAI. The best predictors of TAI were the tract-oriented strain (6–7 %), strain rate (38–40 s\(^{-1})\), and strain times strain rate (1.3–1.8 s\(^{-1})\) values exceeded by 90 % of the brain. These tract-oriented strain and strain rate thresholds for TAI were comparable to those found in isolated axonal stretch studies. Furthermore, we proposed that the higher degree of agreement between tissue distortion aligned with white matter tracts and TAI may be the underlying mechanism responsible for more severe TAI after horizontal and sagittal head rotations in our porcine model of nonimpact TAI than coronal plane rotations.