Protodrilus (Protodrilidae, Annelida) from the southern and southeastern Brazilian coasts

Protodrilus corderoi, Protodrilus ovarium n. sp. and Protodrilus pythonius n. sp. are reported from beaches in southern and southeastern Brazil and described combining live observations with light and electron scanning microscopy studies. Protodrilus corderoi is redescribed from new collections at the type locality, and a neotype for the species is assigned since the original type material no longer exists. New information on reproductive organs, segmental adhesive glands and unpigmented ciliary receptors as well as morphometrics is provided. Protodrilus ovarium n. sp. and P. pythonius n. sp. are formally described. Protodrilus ovarium n. sp. is diagnosed by the presence of separated lateral organs on segments 7–12, three spermioducts of segments 10–12 and salivary glands in segments 1–9. Protodrilus pythonius n. sp. is defined by the presence of separated lateral organs on segments 7–16, long pygidial lobes and body tapering toward the pygidium. The distribution of the different species in more or less spacious habitats seems to be correlated with their gross morphology. Protodrilus pythonius n. sp., with relatively long and wide body and long palps with ciliary bands, was collected in very coarse sandy sediments at a reflective sheltered beach. Conversely, P. corderoi and P. ovarium n. sp., both possessing more slender bodies with shorter, less ciliated palps, occurred in medium-coarse, well-sorted sediments in the more energetic swash zone of exposed intermediate-reflective beaches. The finding of P. pythonius and P. corderoi in nearby beaches corroborates other studies showing a higher morphological variability among species in different habitats within the same geographical area than among species in the same habitat in different geographical areas.     

Neuromuscular structure of the larva to early ancestrula stages of the cyclostome bryozoan Crisia eburnea

For the first time, the development of a cyclostome bryozoan has been studied with immunochemistry and confocal laser scanning microscopy, with emphasis on nerves and muscles. The larva is covered by multiciliated cells, which are latitudinally strongly elongated and show phalloidin-stained cell junctions. We hypothesize that these cells contract at metamorphosis and squeeze the apical invagination and the adhesive sac out. Ectodermal, longitudinal muscle cells extend from the cells of the inner, conical cuticularized part of the apical invagination to the lower part of the corona, around the adhesive sac pore. These muscles are retained in the ancestrula. Scattered monociliated nerve cells are interspersed between the coronal ciliary cells. An equatorial nerve in the larva disappears at metamorphosis. The central, conical part of the cuticle becomes the terminal membrane of the ancestrula, and the underlying ectodermal and mesodermal cell layers differentiate into the polypide bud, forming a deep narrow invagination, differentiating into vestibule–atrium, mouth ring and pharynx–stomach–rectum. Tentacles develop from the ring of cells around the mouth, and a small ganglion with four nerves innervating each of the tentacles develops at the anal side of the mouth. These new findings yield further support for previous homology statements of bryozoan larvae and development.     

The Arabidopsis DDB1 interacting protein WDR55 is required for vegetative development

The CULLIN family of E3 ubiquitin ligases are important regulators of plant development and function. A newly identified class of CULLIN4-RING-E3 ligases (CRL4s) interacts with substrate receptors referred to as DDB1-CUL4 ASSOCIATED FACTORS (DCAFs) via a DDB1 linker protein. We have previously reported that the WD40 protein WDR55 interacts with DDB1A and is thus a putative DCAF. Mutants of WDR55 are embryo lethal, suggesting that a DDB1^WDR55 complex could regulate embryo and endosperm development. Here we report that a weak allele homozygous for wdr55 display pleiotropic phenotypes in the seedling and adult stages, suggesting a novel regulatory role for WDR55 in vegetative development.     

A novel approach to evaluate ELISA antibody coverage of host cell proteins—combining ELISA-based immunocapture and mass spectrometry

Monitoring host cell proteins (HCPs) is one of the most important analytical requirements in production of recombinant biopharmaceuticals to ensure product purity and patient safety. Enzyme-linked immunosorbent assay (ELISA) is the standard method for monitoring HCP clearance. It is important to validate that the critical reagent of an ELISA, the HCP antibody, covers a broad spectrum of the HCPs potentially present in the purified drug substance. Current coverage methods for assessing HCP antibody coverage are based on 2D-Western blot or immunoaffinity-purification combined with 2D gel electrophoresis and have several limitations. In the present study, we present a novel coverage method combining ELISA-based immunocapture with protein identification by liquid chromatography–tandem mass spectrometry (LC–MS/MS): ELISA-MS. ELISA-MS is used to accurately determine HCP coverage of an early process sample by three commercially available anti-Escherichia coli HCP antibodies, evading the limitations of current methods for coverage analysis, and taking advantage of the benefits of MS analysis. The results obtained comprise a list of individual HCPs covered by each HCP antibody. The novel method shows high sensitivity, high reproducibility, and enables tight control of nonspecific binding through inclusion of a species-specific isotype control antibody. We propose that ELISA-MS will be a valuable supplement to existing coverage methods or even a replacement. ELISA-MS will increase the possibility of selecting the best HCP ELISA, thus improving HCP surveillance and resulting in a final HCP profile with the lowest achievable risk. Overall, this will be beneficial to both the pharmaceutical industry and patient safety.