Protamine — antagonist to heparin

Protamine is used for titration of heparin in vitro for diagnosis of hemorrhagic states and for neutralization of heparin in vivo to terminate heparinization. The protamine equivalent varies with the heparin preparation, conditions of testing and, in vivo, with the amount of heparin present in the circulation. The latter depends on time after administration and the hemodynamic and metabolic state of the patient. Protamine, when injected rapidly, will release histamine and agglutinate platelets. Bleeding (spontaneous hemorrhage) demonstrates a multiple breakdown of hemostatic mechanisms due to surgical stress, drugs, exposure of the blood to foreign surfaces, etc. Simple rules for the amount of protamine required for an individual patient based on clinical judgement will be satisfactory in most cases. When hemostasis is not achieved, it must be appreciated that heparin and protamine are only part of a complex deteriorating situation.     

Comparison of spectroscopy technologies for improved monitoring of cell culture processes in miniature bioreactors

Cell culture process development requires the screening of large numbers of cell lines and process conditions. The development of miniature bioreactor systems has increased the throughput of such studies; however, there are limitations with their use. One important constraint is the limited number of offline samples that can be taken compared to those taken for monitoring cultures in large‐scale bioreactors. The small volume of miniature bioreactor cultures (15 mL) is incompatible with the large sample volume (600 µL) required for bioanalysers routinely used. Spectroscopy technologies may be used to resolve this limitation. The purpose of this study was to compare the use of NIR, Raman, and 2D‐fluorescence to measure multiple analytes simultaneously in volumes suitable for daily monitoring of a miniature bioreactor system. A novel design‐of‐experiment approach is described that utilizes previously analyzed cell culture supernatant to assess metabolite concentrations under various conditions while providing optimal coverage of the desired design space. Multivariate data analysis techniques were used to develop predictive models. Model performance was compared to determine which technology is more suitable for this application. 2D‐fluorescence could more accurately measure ammonium concentration (RMSE_CV 0.031 g L^?1) than Raman and NIR. Raman spectroscopy, however, was more robust at measuring lactate and glucose concentrations (RMSE_CV 1.11 and 0.92 g L^?1, respectively) than the other two techniques. The findings suggest that Raman spectroscopy is more suited for this application than NIR and 2D‐fluorescence. The implementation of Raman spectroscopy increases at‐line measuring capabilities, enabling daily monitoring of key cell culture components within miniature bioreactor cultures. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:337–346, 2017     

Can generic knee joint models improve the measurement of osteoarthritic knee kinematics during squatting activity?

Knee joint kinematics derived from multi-body optimisation (MBO) still requires evaluation. The objective of this study was to corroborate model-derived kinematics of osteoarthritic knees obtained using four generic knee joint models used in musculoskeletal modelling – spherical, hinge, degree-of-freedom coupling curves and parallel mechanism – against reference knee kinematics measured by stereo-radiography. Root mean square errors ranged from 0.7° to 23.4° for knee rotations and from 0.6 to 9.0 mm for knee displacements. Model-derived knee kinematics computed from generic knee joint models was inaccurate. Future developments and experiments should improve the reliability of osteoarthritic knee models in MBO and musculoskeletal modelling.     

Colorectal cancer desmoplastic reaction up-regulates collagen synthesis and restricts cancer cell invasion

During cancer cell growth many tumors exhibit various grades of desmoplasia, unorganized production of fibrous or connective tissue, composed mainly of collagen fibers and myofibroblasts. The accumulation of an extracellular matrix (ECM) surrounding tumors directly affects cancer cell proliferation, migration and spread; therefore the study of desmoplasia is of vital importance. Stromal fibroblasts surrounding tumors are activated to myofibroblasts and become the primary producers of ECM during desmoplasia. The composition, density and organization of this ECM accumulation play a major role on the influence desmoplasia has upon tumor cells. In this study, we analyzed desmoplasia in vivo in human colorectal carcinoma tissue, detecting an up-regulation of collagen I, collagen IV and collagen V in human colorectal cancer desmoplastic reaction. These components were then analyzed in vitro co-cultivating colorectal cancer cells (Caco-2 and HCT116) and fibroblasts utilizing various co-culture techniques. Our findings demonstrate that direct cell-cell contact between fibroblasts and colorectal cancer cells evokes an increase in ECM density, composed of unorganized collagens (I, III, IV and V) and proteoglycans (biglycan, fibromodulin, perlecan and versican). The desmoplastic collagen fibers were thick, with an altered orientation, as well as deposited as bundles. This increased ECM density inhibited the migration and invasion of the colorectal tumor cells in both 2D and 3D co-culture systems. Therefore this study sheds light on a possible restricting role desmoplasia could play in colorectal cancer invasion.     

The effect of curcumin in the ectonucleotidases and acetylcholinesterase activities in synaptosomes from the cerebral cortex of cigarette smoke-exposed rats

With the evidence that curcumin may be a potent neuroprotective agent and that cigarette smoke is associated with a decline in the cognitive performance as our bases, we investigated the activities of Ecto‐Nucleoside Triphosphate Diphosphohydrolase (NTPDase), 5'‐nucleotidase and acetylcholinesterase (AChE) in cerebral cortex synaptosomes from cigarette smoke‐exposed rats treated with curcumin (Cur). The experimental procedures entailed two sets of experiments. In the first set, the groups were vehicle, Cur 12·5, 25 and 50 mg·kg^–1; those in the second set were vehicle, smoke, smoke and Cur 12·5, 25 and 50 mg·kg^–1. Curcumin prevented the increased NTPDase, 5'‐nucleotidase and AChE activities caused by smoke exposure. We suggest that treatment with Cur was protective because the decrease of ATP and acetylcholine (ACh) concentrations is responsible for cognitive impairment, and both ATP and ACh have key roles in neurotransmission. Copyright © 2011 John Wiley & Sons, Ltd.     

Differential turnover of isoflavone 7-O-glucoside-6″-O-malonates in Cicer arietinum roots

Pulse labelling experiments and studies with a molecular inhibitor of phenylalanine ammonia lyase showed that in roots of Cicer arietinum formononetin 7-O-glucoside-6″-O-malonate is rapidly metabolized whereas biochanin A 7-O-glucoside-6″-O-malonate appears to be metabolically rather inert.     

Ecology of terrestrial testate amoebae assemblages from coastal Lowlands on Devon Island (NWT,Canadian Arctic)

Summary Fifty-seven taxa belonging to 17 genera of testate rhizopods were recorded in samples of mosses, soils and lichens, collected from the Truelove and Sparbo Hardy Lowlands on the high arctic Devon Island (NWT, Canada). Different groups of habitats are characterized by particular assemblages of Testacea species, with the quantity and quality of the available water as important ecological parameters. The Assulina muscorum-Corythion dubium assemblage characterizes the most dry and acidic moss habitats, while the Paraquadrula irregularis assemblage is to be found in wet and rather neutral habitats. The sedge moss meadows with their more moderately coarse textured and poorly drained soils, bear a Plagiopyxis callida-Plagiopyxis declivis soil-dwelling testate amoebae assemblage. The Centropyxis minuta assemblage lives in the most coarsely textured soils. pH seems to play a subordinate role, not only in the mosses but also in the soils. Multivariate methods (cluster analysis and CANOCO) have been used to separate these different assemblages.     

Conversion of squalene-2(3)epoxide by enzymes of Alnus glutinosa

The incubation of [1-¹⁴C]-squalene-2(3)epoxide ([1-¹⁴C]-SO) with a cell-free extract of Alnus glutinosa gave only cycloartenol in 1% yield.     

Occurrence and accumulation of the granulosis virus of Pieris rapae in treated field plots

The granulosis virus of Pieris rapae, the imported cabbageworm (P. rapae GV), persisted in soil after application. Virus produced by epizootics of the disease in populations of the host accumulated in soil in nontreated plots with concentrations of the virus in soil and on foliage, being similar at harvest to concentrations in virus-treated plots. Also P. rapae GV produced in epizootics of the disease increased concentrations in soil of virus-treated plots and maintained substantial residues of the virus on the foliage. Little virus accumulated in plots treated with some chemical insecticides and Bacillus thuringiensis because few host larvae survived to support late-season epizootics. Small quantities of P. rapae GV were found in heads of cabbage harvested from plots in October, but these residues were not related to plot treatment.A study in which nontreated and virus-treated plots were replanted for 2 years after treatment indicated an increase in concentrations of P. rapae GV in surface soil in years following treatment. Concentrations of P. rapae GV in soil in virus-treated and nontreated plots were similar in the autumn of the year of treatment and in subsequent years of the study. Concentrations were nearly unchanged during the winter, were reduced by cultivation preparatory to planting in the spring, and increased with epizootics of the disease in host populations in summer and autumn.P. rapae GV disease was prevalent during September and October in populations of P. rapae larvae in plots in which substantial concentrations of the virus were found and contributed to late-season control of the pest insect.