A High Performance Gel Filtration Chromatography Method for gamma-Glutamyltransferase Fraction Analysis

The clinical relevance of serum c-glutamyltransferase (GGT) activity, in areas other than hepatic function, has recently been increased by several epidemiological associations. Still, GGT remains a nonspecific test because of the influence of various pathophysiological factors. We devised a procedure based on gel filtration chromatography, followed by postcolumn injection of fluorescent GGT substrate (cglutamyl- 7-amido-4-methylcoumarin), permitting the quantification of GGT fractions in serum or plasma. Plasma GGT molecular weight distribution was analyzed in healthy volunteers (20 males; mean ± SD age 38 ± 10 years; 20 females; age 44 ± 13; total GGT 21 ± 11 for males vs 13 ± 7 for females; P < 0.01). The method is highly sensitive (determination limit: 0.5 U GGT/L), with a linear dynamic range between 0.5 and 150 U/L for each fraction. Four GGT fractions of different molecular weight were detected in all subjects of both genders: b-GGT, m-GGT, s-GGT (likely lipoprotein-bound, molecular masses >2000, 940, and 140 kDa, respectively), and a free fraction (f-GGT, 70 kDa). f-GGT and s-GGT were the main fractions in subjects with lower and higher total GGT activity, respectively. Higher total GGT activity in males is related mainly to f-GGT (P < 0.01). GGT fraction analysis may increase the sensitivity and specificity of the GGT activity test

Different media and supplements modulate the clonogenic and expansion properties of rabbit bone marrow mesenchymal stem cells

<p>Abstract</p> <p>Background -</p> <p>Rabbits provide an excellent model for many animal and human diseases, such as cardiovascular diseases, for the development of new vaccines in wound healing management and in the field of tissue engineering of tendon, cartilage, bone and skin.</p> <p>The study presented herein aims to investigate the biological properties of bone marrow rabbit MSCs cultured in different conditions, in order to provide a basis for their clinical applications in veterinary medicine.</p> <p>Findings -</p> <p>MSCs were isolated from 5 New Zealand rabbits. Fold increase, CFU number, doubling time, differentiation ability and immunophenotype were analyzed.</p> <p>With the plating density of 10 cells/cm²the fold increase was significantly lower with DMEM-20%FCS and MSCs growth was significantly higher with αMEM-hEGF. The highest clonogenic ability was found at 100 cell/cm²with MSCBM and at 10 cell/cm²with M199. Both at 10 and 100 cells/cm², in αMEM medium, the highest CFU increase was obtained by adding bFGF. Supplementing culture media with 10%FCS-10%HS determined a significant increase of CFU.</p> <p>Conclusion -</p> <p>Our data suggest that different progenitor cells with differential sensitivity to media, sera and growth factors exist and the choice of culture conditions has to be carefully considered for MSC management.</p

The Role of Anti-HLA Antibodies in Hematopoietic Stem Cell Transplantation

Donor-specific antihuman leukocyte antigen antibodies (DSHA) have been clearly implicated in graft rejection in solid organ transplantation. Their role in allogeneic hematopoietic stem-cell transplantation (allo-HSCT) remains unclear. We summarize here evidence supporting a role for DSHA in graft failure in animal models of allo-HSCT and in clinical settings whenever no full HLA matching occurs

Effects of recycled materials added to the substrate on nursery trees

Transplant can be a shock for plants, especially when they are moved from the nursery, where the environmental conditions are very favourable, to urbanised areas, where resources and maintenance are scarce. Moreover, some characteristics of urban soils, such alkaline pH and compaction, make it difficult for trees to adapt. Recycled organic material (compost) added as mulch to tree plantings is effective in contrasting compaction in urban soils (Scharenbroch and Watson 2014). Designed soil materials can promote successful urban tree planting, allowing better root growth and plant development (grabosky et al. 2001). The industry of recycled paper produces tons of waste, which in some cases can be a source of pollutants (Tandy et al. 2008). The deinking sludge we choose is a clean material consisting mainly of cellulose, that can be potentially used as a component in plant growth substarte. This material ha a good water retention and porosity and it has been successfully employed as a growing media component (Vannucchi et al. 2017). In the present study the deinking sludge converted into pellets has been added to the growing media of nursery trees, with the aim of increasing the water storage in the substrate and, at the same time, of contrasting soil compaction. In order to get the permission to use the waste in the experiment, an application was submitted to the local planning authorities primarly inteded to manage industrial waste

Use of coarse substrate to increase the rate of water infiltration and the bearing capacity in tree plantings

This work assesses the suitability of recycled crushed bricks and compost, used as a mixed growing substrate, in urban roadside plantings of Tilia cordata Mill. Coarse recycled materials can prevent compaction, improving air and water flows in the rhizosphere, and thus reducing tree mortality or poor growth. In addition, compost is a source of organic matter, contributing to enhance soil quality. Six substrates were tested: 1) soil; 2) crushed bricks 0–30 mm; 3) crushed bricks 6–30 mm; 4) soil + compost (15% v/v); 5) crushed bricks 0–30 mm + compost; and 6) crushed bricks 6–30 mm + compost. Trees were planted in 0.6 × 0.6 × 0.6 m pits, filled with the six different substrates. The water infiltration rate (Kf) and load bearing capacity (Evd) of the substrate were monitored along with the shoot length, and chlorophyll content of the trees. At the end of the trial, the root biomass and total root length were measured. Crushed bricks positively affected Kf, and Evd; the presence of compost decreased the value of bearing capacity. 0–30 crushed brick substrates (with or without compost) increased the linden shoot length and enabled to develop higher root biomass, compared to the soil (+72%) and 6–30 (+37%). The best trade-off between resistance, compaction and plant growth was 0–30 mm crushed bricks. This research provides evidence that a coarse matrix substrate made of crushed bricks is efficient in preventing compaction and allows plant growth

Contraccezione d'emergenza

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THE EIGHTEEN MONTH MORTALITY IS REDUCED WITH A NOVEL IMMUNOSUPPRESSION STRATEGY WHEN AGED LIVERS ARE TRANSPLANTED IN HCV POSITIVE RECIPIENTS

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Pooled human serum: A new culture supplement for bioreactor-based cell therapies. Preliminary results

Background aims: Bone marrow mesenchymal stromal cells (MSCs) are an appealing source for several cell-based therapies. Many bioreactors, such as the Quantum Cell Expansion System, have been developed to generate a large number of MSCs under Good Manufacturing Practice conditions by using human platelet lysate (HPL). Previously, we isolated a novel cell population in human bone marrow, mesodermal progenitor cells (MPCs), which we identified as precursors of MSCs. MPCs may represent an important cell source for regenerative medicine applications. Because HPL gives rise to a homogeneus MSC population, limiting the harvesting of other cell types, we investigated the efficacy of pooled human AB serum (ABS) to provide clinically relevant numbers of both MSCs and MPCs for regenerative medicine applications by using the Quantum System. Methods: Bone marrow aspirates were obtained from healthy adults undergoing routine total hip replacement surgery. The aspirates were used to generate primary cultures in the bioreactor. HPL and ABS were tested as supplements to culture medium. Morphological observations, cytofluorimetric analysis and lactate and glucose level assessments were performed. Results: ABS gave rise to both heterogeneous MSC and MPC populations. About 95% of cells cultured in HPL showed a fibroblast-like morphology and typical mesenchymal surface markers, but MPCs were scarcely represented. Discussion: The use of ABS appeared to sustain large-scale MSC production, as well as promote the recovery of a subset of MPCs, resulting in a suitable alternative to HPL in the cell generation based on the Quantum System