On the identity of Chamaedrilus glandulosus (Michaelsen, 1888) (Clitellata,Enchytraeidae), with the description of a new species

The taxonomy of Chamaedrilus glandulosus (Michaelsen, 1888) s. l., most commonly known previously as Cognettia glandulosa, is revised. A recent molecular systematic study has shown that this taxon harbours two cryptic, but genetically well separated lineages, each warranting species status. In this study these two lineages are scrutinized morphologically, on the basis of Michaelsen’s type material as well as newly collected specimens from Central and Northern Europe. Chamaedrilus glandulosus s. s. is redescribed and Chamaedrilus varisetosus sp. n. is recognized as new to science. The two species are morphologically very similar, differing mainly in size, but seem to prefer different habitats, with Chamaedrilus glandulosus being a larger aquatic species, and Chamaedrilus varisetosus being smaller and mainly found in moist to wet soil.     

IMAC capture of recombinant protein from unclarified mammalian cell feed streams

Fusion‐tag affinity chromatography is a key technique in recombinant protein purification. Current methods for protein recovery from mammalian cells are hampered by the need for feed stream clarification. We have developed a method for direct capture using immobilized metal affinity chromatography (IMAC) of hexahistidine (His6) tagged proteins from unclarified mammalian cell feed streams. The process employs radial flow chromatography with 300–500 μm diameter agarose resin beads that allow free passage of cells but capture His‐tagged proteins from the feed stream; circumventing expensive and cumbersome centrifugation and/or filtration steps. The method is exemplified by Chinese Hamster Ovary (CHO) cell expression and subsequent recovery of recombinant His‐tagged carcinoembryonic antigen (CEA); a heavily glycosylated and clinically relevant protein. Despite operating at a high NaCl concentration necessary for IMAC binding, cells remained over 96% viable after passage through the column with host cell proteases and DNA detected at ～8 U/mL and 2 ng/μL in column flow‐through, respectively. Recovery of His‐tagged CEA from unclarified feed yielded 71% product recovery. This work provides a basis for direct primary capture of fully glycosylated recombinant proteins from unclarified mammalian cell feed streams. Biotechnol. Bioeng. 2016;113: 130–140. © 2015 Wiley Periodicals, Inc.     

Diagnosis of bone and liver metastases in breast cancer comparing tumor markers and imaging techniques

One hundred and forty-seven patients were examined by bone scintigraphy, ultrasonography and scintigraphic scan of the liver, at different times after surgical removal of a breast cancer, to rule out skeletal and hepatic metastases. At the same time as imaging procedures, serum levels of tumor markers (CEA, TPA and CA 15-3) were determined using radioimmunometric methods. One or more markers were elevated in all 13 patients with hepatic metastases; 9 out of 46 patients with bone metastases had all serum markers normal, with a sensitivity of 80%. Combined assay of the markers proved useful, TPA and CA 15-3 showing the best sensitivity in bone metastases, and all three markers in liver metastases.     

Activation of glutathione peroxidase via Nrf1 mediates genistein's protection against oxidative endothelial cell injury

Cellular actions of isoflavones may mediate the beneficial health effects associated with high soy consumption. We have investigated protection by genistein and daidzein against oxidative stress-induced endothelial injury. Genistein but not daidzein protected endothelial cells from damage induced by oxidative stress. This protection was accompanied by decreases in intracellular glutathione levels that could be explained by the generation of glutathionyl conjugates of the oxidised genistein metabolite, 5,7,3',4'-tetrahydroxyisoflavone. Both isoflavones evoked increased protein expression of gamma-glutamylcysteine synthetase-heavy subunit (gamma-GCS-HS) and increased cytosolic accumulation and nuclear translocation of Nrf2. However, only genistein led to increases in the cytosolic accumulation and nuclear translocation of Nrf1 and the increased expression of and activity of glutathione peroxidase. These results suggest that genistein-induced protective effects depend primarily on the activation of glutathione peroxidase mediated by Nrf1 activation, and not on Nrf2 activation or increases in glutathione synthesis.     

The 3D path of body centre of mass during adult human walking on force treadmill

Three-dimensional (3D) path of the body centre of mass (CM) over an entire stride was computed from ground reaction forces during walking at constant average speed on a treadmill mounted on 3D force sensors. Data were obtained from 18 healthy adults at speeds ranging from 0.30 to 1.40 m s^?1, in 0.1 m s^?1 increments. Six subsequent strides were analyzed for each subject and speed (total strides=1296). The test session lasted about 30 min (10 min for walking). The CM path had an upward concave figure-of-eight shape that was highly consistent within and across subjects. Vertical displacement of the CM increased monotonically as a function of walking speed. The forward and particularly lateral displacements of the CM showed a U-shaped relationship to speed. The same held for the total 3D displacement (25.6–16.0 cm, depending on the speed). The results provide normative benchmarks and suggest hypotheses for further physiologic and clinical research. The familiar inverted pendulum model might be expanded to gyroscopic, “spin-and-turn” models. Abnormalities of the 3D path might flag motor impairments and recovery.     

Mild Hypoxia Enhances Proliferation and Multipotency of Human Neural Stem Cells

Background

Neural stem cells (NSCs) represent an optimal tool for studies and therapy of neurodegenerative diseases. We recently established a v-myc immortalized human NSC (IhNSC) line, which retains stem properties comparable to parental cells. Oxygen concentration is one of the most crucial environmental conditions for cell proliferation and differentiation both in vitro and in vivo. In the central nervous system, physiological concentrations of oxygen range from 0.55 to 8% oxygen. In particular, in the in the subventricular zone niche area, it''s estimated to be 2.5 to 3%.

Methodology/Principal Findings

We investigated in vitro the effects of 1, 2.5, 5, and 20% oxygen concentrations on IhNSCs both during proliferation and differentiation. The highest proliferation rate, evaluated through neurosphere formation assay, was obtained at 2.5 and 5% oxygen, while 1% oxygen was most noxious for cell survival. The differentiation assays showed that the percentages of β-tubIII+ or MAP2+ neuronal cells and of GalC+ oligodendrocytes were significantly higher at 2.5% compared with 1, 5, or 20% oxygen at 17 days in vitro. Mild hypoxia (2.5 to 5% oxygen) promoted differentiation into neuro-oligodendroglial progenitors as revealed by the higher percentage of MAP2+/Ki67+ and GalC+/Ki67+ residual proliferating progenitors, and enhanced the yield of GABAergic and slightly of glutamatergic neurons compared to 1% and 20% oxygen where a significant percentage of GFAP+/nestin+ cells were still present at 17 days of differentiation.

Conclusions/Significance

These findings raise the possibility that reduced oxygen levels occurring in neuronal disorders like cerebral ischemia transiently lead to NSC remaining in a state of quiescence. Conversely, mild hypoxia favors NSC proliferation and neuronal and oligodendroglial differentiation, thus providing an important advance and a useful tool for NSC-mediated therapy of ischemic stroke and neurodegenerative diseases like Parkinson''s disease, multiple sclerosis, and Alzheimer''s disease.     

In vitro maturation rates of canine oocytes from anoestrous bitches in simple media

The meiotic competence of canine oocytes collected from anoestrous bitch ovaries and cultured for 72 h in different media was studied. The base culture medium was TCM 199 enriched with 10% fetal bovine serum (TCM); the effect of supplementation with EGF (50 ng x mL(-1)) or ITS (insulin: 10 microg x mL(-1); transferrin: 5.5 microg x mL(-1); selenium: 5 microg x mL(-1)) was also studied. TCM was also compared to a Synthetic Oviductal Fluid (SOF). All the media contained FSH (0.1 UI x mL(-1)), LH (10 UI x mL(-1)), 17beta-oestradiol (4 microg x mL(-1)) and kanamycin. Despite the anoestrous stage of the donor bitches, resumption of meiosis occurred in a high proportion of the oocytes, (mean value 77.3%). The number of oocytes showing the 'germinal vesicle breakdown' nuclear stage was not influenced by the type of the culture medium used. ITS had a positive effect on nuclear progression to later stages (from metaphase I to metaphase II); however, this effect was not statistically significant.     

A Single Amino Acid Change in the Hemagglutinin Protein of Measles Virus Determines Its Ability To Bind CD46 and Reveals Another Receptor on Marmoset B Cells

This paper provides evidence for a measles virus receptor other than CD46 on transformed marmoset and human B cells. We first showed that most tissues of marmosets are missing the SCR1 domain of CD46, which is essential for the binding of Edmonston measles virus, a laboratory strain that has been propagated in Vero monkey kidney cells. In spite of this deletion, the common marmoset was shown to be susceptible to infections by wild-type isolates of measles virus, although they did not support Edmonston measles virus production. As one would expect from these results, measles virus could not be propagated in owl monkey or marmoset kidney cell lines, but surprisingly, both a wild-type isolate (Montefiore 89) and the Edmonston laboratory strain of measles virus grew efficiently in B95-8 marmoset B cells. In addition, antibodies directed against CD46 had no effect on wild-type infections of marmoset B cells and only partially inhibited the replication of the Edmonston laboratory strain in the same cells. A direct binding assay with insect cells expressing the hemagglutinin (H) proteins of either the Edmonston or Montefiore 89 measles virus strains was used to probe the receptors on these B cells. Insect cells expressing Edmonston H but not the wild-type H bound to rodent cells with CD46 on their surface. On the other hand, both the Montefiore 89 H and Edmonston H proteins adhered to marmoset and human B cells. Most wild-type H proteins have asparagine residues at position 481 and can be converted to a CD46-binding phenotype by replacement of the residue with tyrosine. Similarly, the Edmonston H protein did not bind CD46 when its Tyr481 was converted to asparagine. However, this mutation did not affect the ability of Edmonston H to bind marmoset and human B cells. The preceding results provide evidence, through the use of a direct binding assay, that a second receptor for measles virus is present on primate B cells.