Quantitative methods for scoring cell migration and invasion in filter-based assays

The ability of cells to traverse pores in a biocompatible filter provides means for examining cell chemoattraction. Filter-based assays also permit rapid, quantitative assessment of the in vitro migratory and invasive potential of tumor cells. Scoring migration has relied on visual counting of stained cells which appear on the underside of the filter and determining a true percentage score involves arduous counting of cells on both filter surfaces. Visual counting of random fields may be unreliable, and counting all fields is laborious. In the present study we developed and compared two alternative methods for scoring cell numbers in filter-based assays, a colorimetric assay of toluidine blue binding, and a radioassay of cells prelabeled with [3H]thymidine. Each method was evaluated for sensitivity, variability, ease of use and efficiency, and suitability for use in assays of cell migration and invasion. The radiolabeling method proved to be sensitive and reliable and was the most efficient technique. Although less sensitive and specific, the colorimetric dye method offered a rapid and reliable, nonradioactive alternative with the distinct advantage of preserving intact cultures for follow-up visual assessments. We conclude that colorimetric and radiolabel scoring of filter-based assays are reliable and efficient semiautomated methods which provide means to obtain more complete assessments of cell migration and invasion.     

"Cadism": Odd man out.

Comments that L. B. Silverstein's (see record 1992-05293-001) article on child care and maternal employment contributes unfounded and damaging misbeliefs regarding fathers. The dad-as-cad myth suggests intentional malevolence or at least blatant failure by fathers to care for others, and suggests that their failure to share equally in child care and household management is the primary cause of marital conflict. W. B. Johnson and D. Hayes propose a research agenda for fathering that originates in a coherent model of male development in the context of ever-changing environmental demands. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Clinical associations of prenatal ischaemic white matter injury

Neuropathological examinations were carried out at necropsy on 274 cases of intrauterine death or neonatal death at or before three days after birth. Fifty six (20.4%) subjects had evidence of prenatal ischaemic brain damage. On review of the maternal case notes to ascertain antenatal clinical associations there was an increased incidence of intrauterine growth retardation, either based on birth weight for gestational age (odds ratio (OR) 2.0; 95% confidence interval (CI) 1.1 to 3.7) or diagnosed antenatally (OR 2.7; 95% CI 1.3 to 5.6). Oligohydramnios was also more common (OR 2.9; 95% CI 1.2 to 7.0). The association of intrauterine growth retardation and white matter damage remained after excluding fetuses with a major congenital anomaly (OR 2.4; 95% CI 1.1 to 5.1). The findings suggest that chronic intrauterine hypoxia may be associated with damage to cerebral white matter among fetuses and infants who die. The relation between ischaemic white matter damage and cerebral palsy among survivors remains speculative.     

Immobilization of an endopectinlyase on γ-alumina: Study of factors influencing the biocatalytic matrix stability

Endopectinlyase (EC 4.2.2.10) from Aspergillus japonicus was immobilized on to γ-alumina. Adsorption performed at pH 5·0 and a subsequent cross-linking phase using 0·1% glutaraldehyde were the chosen immobilization conditions. The comparison between the main biochemical parameters of the immobilized and free form of the enzyme showed that the immobilization procedure used did not affect the enzyme biochemical properties. The interactions between the carrier and the enzyme are essentially secondary bonding. In fact they depend on the pH and on the presence of phosphate ions in the medium. A tentative chemical model of the biocatalytic matrix thus obtained is proposed.     

Rethinking infant knowledge: toward an adaptive process account of successes and failures in object permanence tasks

Infants seem sensitive to hidden objects in habituation tasks at 3.5 months but fail to retrieve hidden objects until 8 months. The authors first consider principle-based accounts of these successes and failures, in which early successes imply knowledge of principles and failures are attributed to ancillary deficits. One account is that infants younger than 8 months have the object permanence principle but lack means-ends abilities. To test this, 7-month-olds were trained on means-ends behaviors and were tested on retrieval of visible and occluded toys. Means-ends demands were the same, yet infants made more toy-guided retrievals in the visible case. The authors offer an adaptive process account in which knowledge is graded and embedded in specific behavioral processes. Simulation models that learn gradually to represent occluded objects show how this approach can account for success and failure in object permanence tasks without assuming principles and ancillary deficits.     

Monocyte diapedesis through an in vitro vessel wall construct: inhibition with monoclonal antibodies to thrombospondin

Human peripheral blood monocytes were examined for migration across an endothelial cell monolayer in an in vitro vessel wall construct. Few monocytes invaded in the absence of a chemotactic gradient, despite significant adhesion to the endothelial monolayer. However, the addition of zymosan-activated human plasma to the lower compartment, to create a chemotactic gradient across the vessel wall, resulted in significantly enhanced monocyte migration. Pretreatment of the monocytes with monoclonal antibodies to thrombospondin (TSP) dramatically inhibited monocyte diapedesis into the vessel wall. The same treatment inhibited monocyte adhesion to endothelial cells in two-dimensional monolayer cultures as well as in vessel wall constructs (no chemotactic gradient). Of interest, however, the monoclonal antibodies had no inhibitory effect on monocyte migration into collagen gels devoid of endothelial cells in response to the same chemotactic gradient, suggesting the importance of TSP in monocyte-endothelial cell interactions. Monoclonal antibodies to fibronectin and normal mouse immunoglobulin G did not inhibit migration in this model of a vessel wall. Furthermore, monoclonal antibodies to TSP showed no inhibition of human peripheral blood neutrophil migration. Previous studies have shown that monocytes synthesize TSP and express this moiety on their surface. The present data suggest that monocytes may utilize TSP to interact with endothelial cells lining the vessel wall during diapedesis.     

Studie über die Rolle des Vitamin E als Nitritscavenger und N-Nitrosamine Inhibitor

Study on the Role of Vitamin E as Nitrite Scavenger and N-Nitrosamine Inhibitor N-nitrosamine are known for their powerful carcinogenicity. They may be formed under acidic conditions by interaction of nitrosable amino substrates and nitrosating agents such as nitrite, nitrate and nitroso gases. Several of recent investigations demonstrate that certain micronutrients such as vitamin A, C, and E and trace elements selenium and zinc play an important role in cancer prevention. Because of their strong antioxidative, nitrite scavenging and free radical quenching properties, vitamin C and E are capable of inhibiting the peroxidation and nitrosation reaction. This paper summarizes the research work done up to date in the field of inhibition of N-nitrosamine formation by vitamin E. It also includes the latest results of our in vitro study on the inhibitory influence of vitamin E and a mixture of tocopherols as well as the cumulative effect of vitamin C and E on the formation of N-nitroso-N-methylaniline (NMNA). α-tocopherol and the mixture of tocopherols are capable of reducing nitrite concentration in hydrophilic reaction medium and thus inhibits the formation of N-nitrosamine. Both vitamins together have a stronger nitrite scavenger effect than either of them alone. The results of research work done up to date strongly suggest that both vitamins may play an important role in reducing the health hazard of some carcinogenic N-nitroso compounds.     

Agens Faber: Toward a Theory of Artefacts for MAS

Human intelligence has evolved along with the use of more and more sophisticated tools, allowing Homo Faber (from Homo Habilis to Homo Sapiens Sapiens) to cope with environment changes, as well as to adapt the environment to his needs. Analogously, in this seminal paper we introduce the notion of Agens Faber, conveying the idea that agent intelligence should not be considered as separated by the agent ability to perceive and affect the environment—and so, that agent intelligence is strictly related to the artefacts that enable, mediate and govern any agent (intelligent) activity.

Along this line, we first discuss the notion of artefact for MAS in general, then we try to devise out the admissible / required / desirable features of an artefact for MAS. We elaborate on the many sorts of possible relations between agents and artefacts, focusing in particular on the issue of the rational exploitation of artefacts, and also rough out a possible taxonomy of artefacts for MAS.