Supplementary Figure 2 from A Novel Bispecific, Trivalent Antibody Construct for Targeting Pancreatic Carcinoma

PDF file - 718 KB, (A) Release of doxorubicin from milatuzumab (hLL1)-doxorubicin conjugate at pH 5, 37 degrees Celsius and (B) in serum at 37 degrees Celsius

PDF file - 59 KB, Reversed-phase HPLC analyses of cathepsin B incubates and buffer control incubates of ME-CL2E-SN-38, pH 5, 37 degrees Celsius. Values represent % of applied product.

Abstract CD74 is an attractive target for antibody–drug conjugates (ADC), because it internalizes and recycles after antibody binding. CD74 mostly is associated with hematologic tumors but is expressed also in solid cancers. Therefore, ADCs of the humanized anti-CD74 antibody, milatuzumab, were examined for the therapy of CD74-expressing solid tumors. Milatuzumab–doxorubicin and two milatuzumab–SN-38 conjugates with cleavable linkers, differing in their stability in serum and how they release SN-38 in the lysosome, were prepared. CD74 expression was determined by flow cytometry and immunohistology. In vitro cytotoxicity and in vivo therapeutic studies were conducted in the human cancer cell lines A-375 (melanoma), HuH-7 and Hep-G2 (hepatoma), Capan-1 (pancreatic), NCI-N87 (gastric), and Raji Burkitt lymphoma. The milatuzumab–SN-38 ADC was compared with SN-38 ADCs prepared with anti-Trop-2 and anti-CEACAM6 antibodies in xenografts expressing their target antigens. Milatuzumab–doxorubicin was most effective in the lymphoma model, whereas in A-375 and Capan-1 solid tumors, only milatuzumab–SN-38 showed a therapeutic benefit. Despite much lower surface expression of CD74 than Trop-2 or CEACAM6, milatuzumab–SN-38 had similar efficacy in Capan-1 as anti-Trop-2–SN-38, but in NCI-N87, anti-CEACAM6 and anti-Trop-2 conjugates were superior. Studies in two hepatoma lines at a single dose level showed significant benefit over saline controls but not against an irrelevant immunoglobulin G conjugate. CD74 is a suitable target for ADCs in some solid tumor xenografts, with efficacy largely influenced by uniformity of CD74 expression and with SN-38 conjugates providing the best therapeutic responses; SN-38 conjugates were preferable in solid cancers, whereas doxorubicin ADC was better in lymphoma tested. Mol Cancer Ther; 12(6); 968–78. ©2013 AACR.

Supplementary Figure 1 from A Novel Bispecific, Trivalent Antibody Construct for Targeting Pancreatic Carcinoma

The Downton Bone Bed is a Konzentrat-Lagerstätte deposit located in the Welsh Borderlands, United Kingdom. The Downton Bone Bed is late Silurian, considered to be Přídolí in age, and occurs within the Platyschisma Shale Member of the Downton Castle Sandstone Formation. The bone bed is exposed at Weir Quarry (Herefordshire), which this study proposes should become established as the type locality for this stratigraphic horizon, due to the destruction of other localities and lack of access to other sites. As Weir Quarry is one of the last remaining exposures of this unit, the objective of this study is to qualitatively describe the sedimentology, ichnology, and invertebrate palaeontology of the bone bed, to enhance the regional understanding of the palaeogeography, depositional environments and depositional processes of the Welsh Borderlands during the late Silurian. Parasequence thickness and frequency, and sedimentary structures such as hummocky cross stratification, observed within the Downton Castle Sandstone Formation, have traditionally been explained to have formed by sea-level oscillations. These are interpreted to have formed entirely by nearshore to shoreface, shallow marine autogenic sedimentary processes, such as storm events (tempestites) and tidal scour. It is interpreted that formation of the Downton Bone Bed occurred as a by-product of these autogenic sedimentary processes, through winnowing and erosion during storm-driven scour and reworking. The low diversity ichno- and invertebrate fauna observed within the Downton Bone Bed is consistent with a nearshore depositional environment and is indicative of a stressed ecosystem due to fluctuating salinity and oxygen levels.

PDF file - 60 KB, Reversed-phase HPLC analyses of cathepsin B incubates and buffer control incubates of ME-CL2A-SN-38, pH 5, 37 degrees Celsius. Values represent % of applied product.

PDF file - 1.5 MB, Reversed-phase HPLC analyses for cleavage of ME-capped CL2E-SN-38 by cathepsin-B

PDF file - 2 MB, Immunohistological assessment of the distribution of CEACAM6 and Trop-2 in formalin-fixed, paraffin-embedded xenografts of human cancer cell lines

Microplastic is a significant global problem. The rapid screening of environmental matter is highly beneficial to the efficient detection, analysis and mapping of microplastic pollution, however many current laboratory techniques to test samples are time-consuming and often involve hazardous chemicals. SEM-based automated mineralogy (AM) is a uniquely powerful tool for quantifying chemical, mineral and textural properties for a wide-range of sample types. This paper presents an attempt to use AM to identify and quantify microplastic within a heterogeneous surrounding matrix using QEMSCAN ® (Quantification and Evaluation of Minerals by Scanning Electron Microscopy). Here, the standard AM processes are adapted to develop an entirely new methodology, involving the use of a novel mounting medium for sample preparation and the building of a Species Identification Protocol (SIP) using polymer standards. The results show potential, although challenges include the over-quantification of plastic and differentiation from natural matter. Additional challenges relate to limitations regarding the particular AM system used, which places restrictions on methodology, but which may be overcome with newer systems. This study indicates that, with further refinement, AM may have future potential as a high-throughput, cost-effective, initial screening step to identify highly microplastic-polluted areas and accelerate research into establishing solutions.

Abstract Molecular fossils have been used to augment the physical fossil record, charting the expansion of complex life through the Neoproterozoic Era (~1000-541 Ma). This work relies on the hypothesis that C27 steranes preserved in sedimentary rocks originated from cholesterol, the predominant sterol produced by red algae and animals. Following the same logic, C28 and C29 carbon steranes are widely considered to be derived from the sterols of fungi, green algae and some protists. In this study, we demonstrate that the gene 24-C sterol methyltransferase (smt), which is necessary to produce C28 and C29 sterols, is present in segmented worms, an advanced group of animals. Phylogenetic analysis of the relevant gene suggests it was present in the first animals and lost independently in at least seven major lineages. A molecular clock demonstrates that the SMT specific to animals and their closest ancestors was present through the Neoproterozoic. Based on these results, C27 steranes cannot be considered indicative of Neoproterozoic animals, and C28+ steranes are not solely indicative of fungi or green algae. While our results do not necessarily contradict the emerging picture of Neoproterozoic life informed by molecular fossils, they refute the underlying hypothesis that drives the interpretive paradigm.

Eastern Indonesia has a prolonged, complex tectonic history. It is where the Eurasian, Indo-Australian, Caroline and Philippine Sea plates converge, and where processes such as subduction, obduction, slab rollback, rifting, supracrustal extension, lower crustal flow and exhumation are very young or still active.For these reasons, the SE Asia Research Group (SEARG) at Royal Holloway, University of London, has made Eastern Indonesia one of its major research themes in recent years. The SEARG has been conducting geological research in SE Asia since 1982. Work has been undertaken in Indonesia, Malaysia, Thailand, the Philippines, Vietnam and the South China Sea. In 2012 the SEARG is directed by Professor Robert Hall, and involves 12 postgraduate students, 2 postdoctoral researchers, a large number of academic staff, research associates and collaborators in the UK and overseas. The group is funded by a consortium of oil companies.Here we summarise recent and ongoing SEARG projects in Eastern Indonesia. Most of the projects are field-based, but they all also employ new data and techniques, such as 40Ar-39Ar, U-Pb dating (SHRIMP and LA-ICP-MS), Hf isotope dating (LA-MC-ICP-MS), UTh/ He dating, multibeam bathymetry, high quality seismic and remote sensing data.

Earth history is defined by a series of greenhouse and icehouse periods associated with cycles of sea-level rise and fall. Throughout time great oceans such as the Tethys have opened and closed and have seen the beginning of life on Earth. Sea-level variation is driven by a variety of factors including orbital (Milankovitch) cyclicity and the Wilson cycle of the formation and break-up of continents which controls the volume of ocean basins. The volume of water in the oceans is also strongly controlled by the volume of water locked up in ice sheets and glaciers, and is therefore linked to changes in mean global temperatures. Consequently, eustatic sea-level rise and fall occurs synchronously with the retreat and expansion of ice sheets, respectively. Interpretations of sea-level change in geological time have traditionally been made using sequence stratigraphic concepts. However, increasing recognition of the non-uniqueness of sedimentary deposits, particularly through the use of numerical forward models, is challenging this approach. Instead, unequivocal evidence for changes in paleobathymetry must be identified to interpret ancient sea-level change in the rock record. This evidence may come from sea temperature studies using oxygen isotopes as a proxy for sea-level change or from direct evidence for bathymetric change from fossil assemblages that are unique to particular depositional settings. Understanding sea-level change in geological time has applications to modern day climate change research and enables scientists to anticipate the likely response of marine ecosystems to global warming and associated sea-level rise.

The Mississippian Derbyshire and North Wales carbonate platforms were formed in similar tectonic settings within the Pennine and East Irish Sea Basin, respectively. The Derbyshire Platform was surrounded by sub-basins to the north, west and south whilst the North Wales Platform, 130 km west, had a simpler land-attached geometry. Comparison of these age-equivalent platforms allows the controls on sedimentation, at an important juncture in Earth history, to be evaluated. Both platforms are dominated by moderate-to-high-energy, laterally discontinuous facies, with weak evidence for facies cyclicity, suggesting multiple controls on deposition. Influx of siliciclastic mud on the North Wales Platform led to perturbations in carbonate accumulation; along with abundant palaeosols and coal beds this implies a more humid climate, or shallower water depths compared to the Derbyshire Platform. On both platforms, exposure surfaces can rarely be correlated over >500 m except for a regionally correlative palaeokarstic surface at the Asbian–Brigantian boundary. This exposure event appears to coincide with a significant regional facies change. Given the lack of evidence for ordering and cyclicity within the strata, the Asbian–Brigantian boundary may mark a significant event that could reflect the onset of a transitional climate, prior to the second glaciation event in the Late Palaeozoic Ice Age.

The New Guinea Limestone Group was deposited across much of New Guinea, including the Indonesian provinces of West Papua and Papua, as part of a widespread shallow-water carbonate platform during the Paleogene and Neogene. This platform was drowned beneath deeper-water strata from the Middle to Late Miocene. Review of biostratigraphic and seismic data from the Aru Basin, offshore New Guinea, reveals a drowning succession c. 600 m thick deposited during a drowning event that lasted around 4 Ma. The objective of this study was to create a well-to-seismic tie from a single well in the study area using biostratigraphic, seismic and log data. The well-to-seismic tie was built to constrain a new velocity model to better image the drowned carbonate platform and understand the reservoir potential of the drowning succession in the zone of interest using two complimentary techniques: seismic reservoir characterization and numerical stratigraphic forward modelling. The well-to-seismic tie was achieved by matching significant biostratigraphic events, such as unconformities, with seismic horizons using stratigraphy-to-seismic. Modern stratigraphic and seismic reservoir characterization techniques, including stratigraphy-to-seismic, numerical forward modelling, velocity model building, rock physics and seismic inversion, were applied to predict rock properties such as lithology and porosity within the drowning succession.

Papua New Guinea resides in a complex tectonic junction between the Australian continent, the Southwest Pacific, and Southeast Asia. Resolving the plate tectonic evolution of this region has proven difficult to date due to the plethora of contradicting geologic models that stem from a lack of constraining regional datasets. A growing body of geochronologic, geochemical and isotopic data acquired over the past decade, combined with new field observations, has led to a progressive shift from a largely autochthonous terrane model, to one of allochthonous terranes and accretion at the northern Australian continental margin. Findings from a compilation of zircon and biostratigraphic age data throughout the Papuan Peninsula and the New Guinea Fold and Thrust, and Mobile Belts, extending west to the Bird’s Head of West Papua, provide a robust evidence-based provenance model for the origin of these terranes. The evaluation of inherited and detrital zircon age populations indicates that many of the terranes are allochthonous in nature and that existing tectonic reconstructions require major revision. For example, a reconstruction that infers a North Queensland provenance for clastic rocks of the Papuan Peninsula based on closure of the Coral Sea is no longer supported by the available data. Instead, some of the allochthonous terranes are now interpreted to be derived from eastern Australia, with provenance of the Papuan Peninsula more akin to that of New Caledonia than North Queensland.

Parasequence thickness and frequency are traditionally interpreted to be controlled by allocyclic processes such as oscillations in eustatic sea-level. However, the use of numerical forward models is challenging these concepts. Outcrop data from Ras Al-Khaimah (UAE) were incorporated into numerical forward models and used to replicate parasequences from the Upper Kharaib Reservoir Unit. Results indicate that clinoform geometries within the parasequences can form by autocyclic, rather than allocyclic, processes. Stratigraphic, sedimentological and palaeoenvironmental interpretations made from outcrops of Upper Kharaib carbonate clinoform parasequences at Wadi Rahabah, Ras Al-Khaimah, were used to build a numerical stratigraphic forward model. Numerical stratigraphic forward models produce fully quantitative three-dimensional deterministic models that replicate and predict the spatial distribution of stratal geometries, stacking patterns, sedimentary thickness and facies formed under a set of predefined input parameters and boundary conditions. A CarboCAT numerical model of carbonate deposystems that uses cellular automata to determine the distribution and lithofacies of heterogeneous carbonate strata in three dimensions (Burgess, 2013) was used to replicate the Upper Kharaib parasequence geometries. Results of the numerical forward model show that carbonate clinoform parasequences from the Upper Kharaib Reservoir Unit can be generated by an autocyclic Ginsburg-type mechanism of sediment transport and shoreline progradation (e.g. Burgess and Wright, 2003; Burgess, 2006). Ginsburg (1971) proposed that progradational, regressive, cycles in carbonates can be formed independently of external eustatic sea-level or tectonically driven forces by autocyclic processes including wind-driven or tidal sediment transport. As sediment is transported towards the shore by autocyclic processes, seaward progradation of the sediment wedge occurs forming a regressive cycle. Alternating transgressive-regressive autocycles are deposited as production rates are less than or exceed the rate of subsidence, respectively. CarboCAT models indicate that the degree of clinoform progradation within the Upper Kharaib is controlled by the rate of relative sea-level rise, driven by localised subsidence, and sediment transport. Thick aggradational parasequences are produced where the rate of relative sea-level rise is the primary control on the deposition of the clinoforms. Parasequence frequency increases where the rate of sediment transport exerts a stronger control than the rate of relative sea-level rise on the deposition of the clinoforms. Observations from numerical forward models have implications for the distribution of reservoir intervals within the Upper Kharaib. Parasequences formed by autocyclic process produce heterogeneous reservoirs with complex facies mosaics. Lateral heterogeneity and variable thicknesses within these reservoirs is more difficult to correlate and trace across fields than simple stacked, layer cake, parasequences created by sea-level oscillations.

This paper presents a biostratigraphic study of upper Miocene to lower Pliocene sedimentary successions from SE Romania based on palynological and micropalaeontological analyses integrated with geophysical wireline log profiles. The data derive from five wells drilled in an area of the Dacian Basin where no previous biostratigraphic data are available. A detailed correlative framework of palynological datums for the studied interval is proposed, which complements and supplements the ostracod and mollusc-based biostratigraphy traditionally used for age determination in the Neogene of the Dacian Basin. Interpretation of the microfossil assemblages in terms of depositional environments enables correlation of the recorded bioevents with regional palaeoenvironmental changes recognised in more marginal areas of the Dacian Basin. Two major marine incursions into the basin are represented by influxes of the benthonic foraminifer Ammonia beccarii and used to define the Khersonian - Meotian and the Mcotian Pontian boundaries. Between these two transgressions, the Meotian is interpreted as a period with at least a certain degree of marine influence. The occurrence of dinocyst taxa usually found in Mediterranean and Atlantic records suggests a connection between Paratethys and the Mediterranean, which probably occurred via the Black Sea. After the Meotian-Pontian marine flooding, a diverse endemic microflora derived from the Pannonian Basin is first recorded in the lower Pontian succession. The signature of a reduction in water level during the middle Pontian within the study area is interpreted to be represented in the successions by an interval dominated by distinctive sand units. The endemic Pannonian dinocysts experienced a significant increase in abundance in the late Pontian, indicating the development of brackish conditions and a water level rise during this period. Due to the absence of foraminifera, or any other indicator of marine conditions, it is probable that this water level rise was related to a positive hydrological balance, as suggested in the Focsani region. The Pontian-Dacian boundary is determined by a transition from brackish to fresh depositional environments, which is associated with a change in the microfloras and the first occurrence of coal layers.