A Porous Media Approach to Finite Deformation Behaviour in Soft Tissues

The present work presents a porous medium formulation for the biomechanical analysis of soft tissues. An updated Lagrangian approach is developed to study the coupled effects of low speed flows of fluid phases, in partially or fully saturated conditions, and the finite deformation occurring in the solid matrix. The procedure developed allows both for the evaluation of coupled geometric and material non-linearities. The main theoretical and computational aspects of this multiphase formulation are discussed. The finite element method is used for the numerical solution of the resulting coupled system of equations. A reference case is reported with regard to healthy and degenerative phases of intervertebral segment. Results reported allow for a detailed interpretation of the formulation reliability, also by comparison with existing experimental data. In particular, the role played by the fluid on the load carrying mechanism is pointed out, thus stressing the importance of a multiphase approach to the overall behaviour of the spinal motion segment in time.     

Numerical analysis of titanium cast devices for dental implantology

The present work pertains to a numerical investigation of the casting process of titanium devices adopted for dental implantology. The analysis of the titanium framework that connects different abutments, in a multi-implant configuration, is performed evaluating the characteristics of the material that depend on the manufacturing procedure. The connecting bar is obtained by a foundry process that is controlled through a numerical simulation by using a control volume technique. This analysis leads to the possibility to have detailed information on the process and control of the quality of the microstructure of the material produced that proves to be highly beneficial for defining mechanical properties. On the basis of the results obtained, a subsequent stress analysis can be performed, addressed to highlight critical conditions. Functional response of the whole bar-implant framework is studied by means of a numerical model, based on the geometric element method. This technique is particularly suited for describing complex morphology of the implant site. The present approach addresses a higher quality definition of the reliability of the device used in dental practice and represents a valuable tool in assisting optimisation procedures pertaining to manufacturing. In fact, the improvement of titanium devices is intended not only with regard to their mechanical performances, but also to requirements pertaining to manufacturing and clinical practice.     

Acceptor specificity of the human leukocyte alpha3 fucosyltransferase: role of FucT-VII in the generation of selectin ligands

The alpha3 fucosyltransferase, FucT-VII, is one of the key glycosyltransferases involved in the biosynthesis of the sialyl Lewis X (sLex) antigen on human leukocytes. The sialyl Lewis X antigen (NeuAcalpha(2-3)Galbeta(1-4)[Fucalpha(1-3)]GlcNAc-R) is an essential component of the recruitment of leukocytes to sites of inflammation, mediating the primary interaction between circulating leukocytes and activated endothelium. In order to characterize the enzymatic properties of the leukocyte alpha3 fucosyltransferase FucT-VII, the enzyme has been expressed in Trichoplusia ni insect cells. The enzyme is capable of synthesizing both sLexand sialyl-dimeric-Lexstructures in vitro , from 3'-sialyl-lacNAc and VIM-2 structures, respectively, with only low levels of fucose transfer observed to neutral or 3'-sulfated acceptors. Studies using fucosylated NeuAcalpha(2-3)-(Galbeta(1- 4)GlcNAc)3-Me acceptors demonstrate that FucT-VII is able to synthesize both di-fucosylated and tri-fucosylated structures from mono- fucosylated precursors, but preferentially fucosylates the distal GlcNAc within a polylactosamine chain. Furthermore, the rate of fucosylation of the internal GlcNAc residues is reduced once fucose has been added to the distal GlcNAc. These results indicate that FucT-VII is capable of generating complex selectin ligands, in vitro , however the order of fucose addition to the lactosamine chain affects the rate of selectin ligand synthesis.      

Immunotoxins to the HER-2 oncogene product: Functional and ultrastructural analysis of their cytotoxic activity

Two immunotoxins were prepared using monoclonal antibodies (mAb) directed towards two distinct epitopes of the gp185^HER-2 extracellular domain, and the type I ribosome inactivating protein (RIP) plant toxin saporin 6. Cell protein synthesis inhibition assay reveals that the immunotoxins display a potent and specific cytotoxicity that is characterized by a slow rate, since the time required to inhibit incorporation of radiolabeled leucine completely ranges from 36 h to 60 h depending on the target cell line and the immunotoxin. Because this feature may hamper the immunotherapeutic use of these conjugates we analysed this further by studying the early phases of internalization of immunotoxins by immunoelectron microscopy. The results of this study have demonstrated that the distribution pattern of the immunotoxins and of the unconjugated mAb over the cell surface overlaps. Similarly the mAb and immunotoxins are internalized into the cell by two different pathways: via clathrin-coated pits or via smaller uncoated pits and vesicles. A higher degree of internalization is achieved when the two immunotoxins are used in combination. Unlike the slow kinetics of cell intoxication the process of immunotoxin endocytosis is characterized by a rapid rate of internalization (above 40% at 5 min in the SK-BR-3 cell line). Although these findings provide no clue to explain the mechanisms of the slow rate of cytotoxicity of the two immunotoxins their rapid internalization indicates that these reagents can be exploited in immunotherapeutic approches to gp185^HER-2-expressing malignancies.     

Abrupt permafrost thaw drives spatially heterogeneous soil moisture and carbon dioxide fluxes in upland tundra

Permafrost thaw causes the seasonally thawed active layer to deepen, causing the Arctic to shift toward carbon release as soil organic matter becomes susceptible to decomposition. Ground subsidence initiated by ice loss can cause these soils to collapse abruptly, rapidly shifting soil moisture as microtopography changes and also accelerating carbon and nutrient mobilization. The uncertainty of soil moisture trajectories during thaw makes it difficult to predict the role of abrupt thaw in suppressing or exacerbating carbon losses. In this study, we investigated the role of shifting soil moisture conditions on carbon dioxide fluxes during a 13-year permafrost warming experiment that exhibited abrupt thaw. Warming deepened the active layer differentially across treatments, leading to variable rates of subsidence and formation of thermokarst depressions. In turn, differential subsidence caused a gradient of moisture conditions, with some plots becoming consistently inundated with water within thermokarst depressions and others exhibiting generally dry, but more variable soil moisture conditions outside of thermokarst depressions. Experimentally induced permafrost thaw initially drove increasing rates of growing season gross primary productivity (GPP), ecosystem respiration (R_eco), and net ecosystem exchange (NEE) (higher carbon uptake), but the formation of thermokarst depressions began to reverse this trend with a high level of spatial heterogeneity. Plots that subsided at the slowest rate stayed relatively dry and supported higher CO₂ fluxes throughout the 13-year experiment, while plots that subsided very rapidly into the center of a thermokarst feature became consistently wet and experienced a rapid decline in growing season GPP, R_eco, and NEE (lower carbon uptake or carbon release). These findings indicate that Earth system models, which do not simulate subsidence and often predict drier active layer conditions, likely overestimate net growing season carbon uptake in abruptly thawing landscapes.     

Nuclear DNA changes within Helianthus annum L.: changes within single progenies and their relationships with plant development

Summary The variations in the basic nuclear DNA content, which previous results indicated to occur within one and the same progeny of Helianthus annuus, were studied in detail and correlated with certain developmental features of the plants. The size and organization of the genome of seedlings obtained from seeds (achenes) collected at the periphery (P-seedlings) or in the middle (M-seedlings) of the flowering heads of plants belonging to a line selfed for 10 years were compared. Cytophotometric determinations indicated that the nuclear DNA content of P-seedlings is 14.7% higher than that of M-seedlings. Thermal denaturation and reassociation kinetics of extracted DNAs showed that variations in the redundancy of repetitive DNA, in particular of a family of medium repeated sequences with a Cot range of 2–100, account for the differences in genome size. These findings were confirmed by the results of molecular hybridizations (slot blots), which also indicated a higher amount of ribosomal DNA in the P-seedlings than in the M-seedlings. Cell proliferation is affected by DNA content, and mitotic cycle time is 1h30 longer in the P-seedlings. By studying mature plants, positive correlations were also found between genome size and both the surface area of leaf epidermal cells (P0.01) and flowering time (P0.001). It is suggested that the variations of nuclear DNA content and organization observed play a role in determining developmental variability in plant populations, which may be of importance in buffering the effects of changing environmental conditions.     

The effect of salts on the kinetics of iron release from N-terminal and C-terminal monoferrictransferrins

The kinetics of iron release from N-terminal and C-terminal monoferric human transferrins has been studied using EDTA as the accepting chelate. In the absence of added salts iron release from the N-terminal site is more facile but the relative lability can be reversed by the addition of NaClO₄, NaCl and LiCl. The results indicate that both anions and cations can affect the lability of the two sites. Since the relative lability of the two monoferrictransferrins is affected by fairly moderate concentrations of NaCl and NaClO₄ we suggest that the ionic composition serum may play an important role in determining the observed distribution of iron among the sites. A new method for the preparation of N-terminal monoferrictransferrin is described.     

Ontogeny of human Ia antigens

Indirect immunofluorescence (IIP) staining of tissues from human fetuses (ages ranging from 8 to 32 weeks of intrauterine life) with monoclonal antibodies (MoAb) to monomorphic determinants of Ia antigens and HLA-A,B,C antigens has shown that both types of antigens are already detectable in tissues of 8-week-old fetuses. Ia antigens and HLA-A,B,C antigens reach their almost-complete tissue distribution after 32 and 24 weeks of intrauterine life, respectively. The structure of Ia antigens synthesized by fetal thymus cells is similar to that of B-lymphoid cell-derived Ia antigens. Ia antigen-bearing thymic fetal cells can stimulate allogeneic lymphocytes in mixed lymphocyte reactions (MLRs). These reactions are blocked by monoclonal antibodies to monomorphic determinants of human Ia antigens and of HLA-A,B, antigens.