A comparison of peripheral blood stem cell collection outcomes for multiple myeloma; mobilization matters in the era of IMiD induction.

Collection of peripheral blood stem cells (PBSCs) for autologous stem cell transplant (ASCT) requires mobilization from the bone marrow. There is variation in mobilization choice; during the COVID-19 pandemic BSBMT&CT guidelines recommended using granulocyte-colony stimulating factor (G-CSF) alone to minimize the use of chemotherapy. We report on the impact of mobilization regimen on stem cell collection, and whether IMiD-containing induction therapy impacts on mobilization and consequently transplant engraftment times for 83 patients undergoing ASCT at Leeds Teaching Hospitals. Cyclophosphamide plus G-CSF (cyclo-G) mobilization yielded more CD34+ cells (8.94 vs. 4.88 ×106/kg, p = < 0.0001) over fewer days (1.6 vs. 2.4 days, p = 0.007), and required fewer doses of salvage Plerixafor than G-CSF only (13.6% vs. 35%, p = 0.0407). IMiD-containing induction impaired all of these factors. CD34+ doses > 8×106/kg were more frequent with Cyclo-G (62% vs. 11%, p = 0.0001), including for those receiving IMiD 1st line induction (50% vs. 13.3%, p = 0.0381). Note that 92.6% of those receiving IMiD-free inductions were mobilized with Cyclo-G. The novel agents used in modern induction regimens (e.g Daratumumab) have been shown to impair yields, increasing the importance of optimizing mobilization regimens in the first instance. Furthermore, as cellular therapies become established in the management of multiple myeloma emerging data highlights the potential benefits of stem cell top up in the management of the haematological toxicities of these therapies. Our findings support re-adoption of Cyclo-G as the gold standard for mobilization to optimize PBSC harvesting and ensure sufficient cells for subsequent ASCTs.

'Lophenteropneust' hypothesis refuted by collection and photos of new deep-sea hemichordates.

The deep ocean is home to a group of broad-collared hemichordates--the so-called 'lophenteropneusts'--that have been photographed gliding on the sea floor but have not previously been collected. It has been claimed that these worms have collar tentacles and blend morphological features of the two main hemichordate body plans, namely the tentacle-less enteropneusts and the tentacle-bearing pterobranchs. Consequently, lophenteropneusts have been invoked as missing links to suggest that the former evolved into the latter. The most significant aspect of the lophenteropneust hypothesis is its prediction that the fundamental body plan within a basal phylum of deuterostomes was enteropneust-like. The assumption of such an ancestral state influences ideas about the evolution of the vertebrates from the invertebrates. Here we report on the first collected specimen of a broad-collared, deep-sea enteropneust and describe it as a new family, genus and species. The collar, although disproportionately broad, lacks tentacles. In addition, we find no evidence of tentacles in the available deep-sea photographs (published and unpublished) of broad-collared enteropneusts, including those formerly designated as lophenteropneusts. Thus, the lophenteropneust hypothesis was based on misinterpretation of deep-sea photographs of low quality and should no longer be used to support the idea that the enteropneust body plan is basal within the phylum Hemichordata.

A new species of the remarkable brittle star genus Astrophiura (Echinodermata: Ophiuroidea) from the western Atlantic Ocean.

Astrophiura caroleae, new species, is described from off Curacao in the southern Caribbean, and from the western Gulf of Mexico, in depths of 244 to 434 meters. This new species, the first in the genus Astrophiura to be described from the Atlantic Ocean, has a distinctive combination of characters, including regularly arranged primary plates, large radial shields whose radial edges are in contact for their entire visible length, and prominent tubercles on central and radial plates. The mottled reddish coloration of the dorsal surface of this species usually contrasts with the color of the substratum, rendering it readily visible in situ, despite its disc diameter of less than 10 mm. Like its congeners, A. caroleae is gonochoric, the gonads of females containing conspicuous masses of bright orange eggs that are approximately 165 µm in diameter. DNA Barcoding data are provided for this new species, these are the first for Astrophiura.

Euryalid brittle stars from the International Indian Ocean Expedition 1963-64 (Echinodermata: Ophiuroidea: Euryalida).

Fourteen species of euryalids are reported from collections made in the eastern and western Indian Ocean during the International Indian Ocean Expedition. Three new species are described: Astrocladus goodingi from the Comoro Islands, Astrocladus socotrana from the northwestern Indian Ocean near the island of Socotra, and Astrocyclus somaliensis from off northeastern Somalia. Range extensions are documented for Asteronyx luzonicus, Astroglymma sculptum, Astrocladus ludwigi, Asteromorpha capensis and Squamophis amamiensis.

Megafauna of the UKSRL exploration contract area and eastern Clarion-Clipperton Zone in the Pacific Ocean: Echinodermata.

BACKGROUND: There is growing interest in mining polymetallic nodules from the abyssal Clarion-Clipperton Zone (CCZ) in the tropical Pacific Ocean. Despite being the focus of environmental studies for decades, the benthic megafauna of the CCZ remain poorly known. In order to predict and manage the environmental impacts of mining in the CCZ, baseline knowledge of the megafauna is essential. The ABYSSLINE Project has conducted benthic biological baseline surveys in the UK Seabed Resources Ltd polymetallic-nodule exploration contract area (UK-1). Prior to these research cruises in 2013 and 2015, no biological studies had been done in this area of the eastern CCZ. NEW INFORMATION: Using a Remotely Operated Vehicle and Autonomous Underwater Vehicle, the megafauna within the UKSRL exploration contract area (UK-1) and at a site ~250 km east of the UK-1 area were surveyed, allowing us to make the first estimates of megafaunal morphospecies richness from the imagery collected. Here, we present an atlas of the abyssal echinoderm megafauna observed and collected during the ABYSSLINE cruises to the UK-1 polymetallic-nodule exploration contract area in the CCZ. There appear to be at least 62 distinct morphospecies (13 Asteroidea, 5 Crinoidea, 9 Echinoidea, 29 Holothuroidea and 6 Ophiuroidea) identified mostly by imagery but also using molecular barcoding for a limited number of animals that were collected. This atlas will aid the synthesis of megafaunal presence/absence data collected by contractors, scientists and other stakeholders undertaking work in the CCZ, ultimately helping to decipher the biogeography of the megafauna in this threatened habitat.

Examining the feasibility of clinical grade CD271+ enrichment of mesenchymal stromal cells for bone regeneration.

INTRODUCTION: Current clinical trials utilize mesenchymal stromal cells (MSCs) expanded in culture, however these interventions carry considerable costs and concerns pertaining to culture-induced losses of potency. This study assessed the feasibility of new clinical-grade technology to obtain uncultured MSC isolates from three human intra-osseous tissue sources based on immunomagnetic selection for CD271-positive cells. MATERIALS AND METHODS: MSCs were isolated from bone marrow (BM) aspirates or surgical waste materials; enzymatically digested femoral heads (FHs) and reamer irrigator aspirator (RIA) waste fluids. Flow cytometry for the CD45-/lowCD73+CD271+ phenotype was used to evaluate uncultured MSCs before and after selection, and to measure MSC enrichment in parallel to colony forming-unit fibroblast assay. Trilineage differentiation assays and quantitative polymerase chain-reaction for key transcripts involved in bone regeneration was used to assess the functional utility of isolated cells for bone repair. RESULTS: Uncultured CD45-/lowCD271+ MSCs uniformly expressed CD73, CD90 and CD105 but showed variable expression of MSCA-1 and SUSD2 (BM>RIA>FH). MSCs were enriched over 150-fold from BM aspirates and RIA fluids, whereas the highest MSC purities were obtained from FH digests. Enriched fractions expressed increased levels of BMP-2, COL1A2, VEGFC, SPARC and CXCL12 transcripts (BM>RIA>FH), with the highest up-regulation detected for CXCL12 in BM (>1300-fold). Following culture expansion, CD271-selected MSCS were tri-potential and phenotypically identical to plastic adherence-selected MSCs. DISCUSSION: A CD271-based GMP-compliant immunomagnetic selection resulted in a substantial increase in MSC purity and elevated expression of transcripts involved in bone formation, vascularisation and chemo-attraction. Although this technology, particularly from RIA fluids, can be immediately applied by orthopaedic surgeons as autologous therapy, further improvements in MSC purities and pre-clinical testing of product safety would be required to develop this process for allogeneic applications.

A new species of Western Atlantic sea lily in the family Bathycrinidae (Echinodermata: Crinoidea), with a discussion of relationships between crinoids with xenomorphic stalks.

A new species in the family Bathycrinidae is described from abyssal depths from the Bahamas. It is referred to the recently established genus Discolocrinus, which formerly comprised a single species D. thieli Mironov, 2008 from the Eastern Pacific. Discolocrinus iselini n. sp. is characterized by large body size, high tegmen with tube-like upper region, extremely elongated IBr1 and IBr2, large knobby processes on primibrachials, and overgrowth of soft tissue on the pinnules, the tissue containing numerous perforated or imperforate ossicles of varying size and form. Differences between Discolocrinus and other bathycrinids may seem to be of taxonomic importance at the family level, but knowledge of the morphology and variability of both species of Discolocrinus is incomplete and, until a richer material becomes available, the genus should remain in family Bathycrinidae. Representatives of five families with xenomorphic stalks were examined to characterize the genera on the basis of number or form of knobby processes. These processes occur in two families with differing external morphology: ten-armed Bathycrinidae and five-armed Bourgueticrinidae. They also occur in the comatulid family Atelecrinidae. This similarity might seem to indicate a close relationship between the three families. However, morphological analysis supports the separation of the families Caledonicrinidae and Septocrinidae from Bathycrinidae despite the fact that they share a xenomorphic stalk and IBr2ax. These conclusions are in agreement with results of recent molecular studies.

Clinical grade OK432-activated dendritic cells: in vitro characterization and tracking during intralymphatic delivery.

Dendritic cells (DC) are under intense preclinical and early clinical evaluation for the immunotherapy of cancer. However, the optimal culture conditions and route of delivery for DC vaccination have not been established. Here we describe the first human application of DC matured with the bacterial agent OK432 (OK-DC), using a short-term serum-free culture protocol, which generates mature DC from CD14+ precursors after 5 days. These cells were prepared within the framework of a National Blood Service facility, demonstrating that DC represent a product which is potentially deliverable alongside current standardized cell therapies within the UK National Health Service. In vitro analysis confirmed that OK-DC were mature, secreted tumor necrosis factor-alpha, interleukin-6, and interleukin-12, and stimulated both T cell and natural killer cell function. To explore effective delivery of OK-DC to lymph nodes, we performed an initial clinical tracking study of radioactively labeled, unpulsed OK-DC after intralymphatic injection into the dorsum of the foot. We showed that injected DC rapidly localized to ipsilateral pelvic lymph nodes, but did not disseminate to more distant nodes over a 48-hour period. There was no significant toxicity associated with OK-DC delivery. These results show that OK-DC are suitable for clinical use, and that intralymphatic delivery is feasible for localizing cells to sites where optimal priming of innate and adaptive antitumor immunity is likely to occur.