Blood and marrow transplantation (BMT) has become an increasingly acceptable treatment for children with life threatening malignant diseases. Survival rates for transplant recipients vary from 23% to 63%. Children with complications from BMT, typically die in the hospital after a prolonged stay. The parental decision to allow a child to die a natural death is typically made in an aura of emotional duress and bewilderment at the complexity and volume of new information that must be assimilated.The purpose of this study was to describe the process of parental decision making for Do Not Resuscitate (DNR) or to withdraw life support in pediatric BMT.The framework for this study was developed from the author's epistemology that blends neo-modernism (recognition of individual uniqueness yet acknowledgment that certain underlying universal principals exist) with the idea that the nature of all things may be viewed as an ongoing, self-constructing process.Grounded theory methodology was used. The sample (determined through theoretical sampling) consisted of seven parents of children who died following BMT and for whom the parent made an end-of-life decision. Data was analyzed using constant comparative analysis, a method that combines both substantive and theoretical coding of data with a qualitative style of theory development.The realization of parental knowing was the process that parents used to navigate the human problem of having to make the end-of-life decision for their children who were dying following blood and marrow transplantation. This process consisted of four categories: Developing Trust, Committed to Seeing It Through, Facing My Worst Fear, and Acceptance of Self.The knowledge gained from this study will inform nurses who care for children who are dying following pediatric BMT. Strategies may be developed that will assist nurses to support the development of parental trust, to help sustain the commitment of parents as they move through the BMT treatment journey, and to assist parents as they face their worst fear. As a result, parents should be better able to achieve an acceptance for themselves that will facilitate a more satisfying experience of the ever changing process occurring in their own lives.

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Bone marrow transplantation (BMT) involves the use of maximal doses of chemotherapy and total body irradiation. As a result, even well-nourished patients exhibit negative nitrogen balance and hypoproteinemia in the post-transplant period, despite a high energy and protein intake from Total Parenteral Nutrition (TPN). The purpose of this research was to investigate the impact of cytotoxic therapy, with a view toward explaining and improving the response to nutritional support. Stool, urine and serum biochemistry were studied prospectively in 10 BMT patients. Analysis of stool revealed increased sodium and decreased potassium. Examination of serum electrolytes indicated hyponatremia and hyperkalemia. A significant decrease in nitrogen balance, serum albumin and net protein utilization immediately followed the disturbances in serum electrolytes; improvement began as serum sodium and potassium returned to normal. Thus, electrolyte imbalance may have reduced the capacity of cells to utilize nitrogen. Lowering the volume of TPN dramatically decreased serum electrolyte aberrations and improved nitrogen utilization.

The alteration of insulin-like growth factor I (IGF-I) and its binding proteins (IGFBP) and their effects on proliferation and differentiation of murine bone marrow-derived macrophage (BMM) precursors were investigated. Bone marrow cells exposed to 20% L929-fibroblast conditioned medium (LCM) were cultured in serum-free medium for 24 h. Western ligand blotting (WLB) analysis detected four bands in all samples. 41-kDa and 30-kDa bands were detected after 12 h and remained constant during BMM differentiation. The 28-kDa and 25-kDa proteins were almost undetectable until day 2, but accumulated significantly from day 3 to day 7. Immunoblotting analysis verified these two bands as IGFBP-4. Northern blotting analyses detected both IGFBP-4 and IGFBP-3 mRNA in the cells. The 41-kDa protein was postulated to be IGFBP-3 in a glycosylated form. The identity of the 30-kDa band is not known. Northern blotting analysis showed that IGF-I mRNA level increased in a time-dependent manner until day 3, and decreased thereafter during BMM differentiation. The effect of IGF-I and its analogs on cell proliferation was studied by ($ sp3$H) thymidine incorporation. IGF-I and its analogs enhanced cell proliferation of freshly isolated bone marrow cells. Both IGF-I and long R$ sp3$ IGF-I, but not des(1-3)IGF-I, continued to exert a stimulating effect on day 1, although to a lesser extent. The effect of IGF-I and its analogs on BMM differentiation was studied by checking morphology, non-specific esterase-1 (NSE-1) activity, and mannose receptor expression. No significant differences in morphology and NSE-1 activity were observed among the treatment groups. There was no difference of mannose receptor expression on day 4 between the IGF-I group and the control cells, whereas long R${ sp3}$ and des(1-3)IGF-I increased the receptor number by 260% and 228% respectively, with less increased K$ rm sb{d}$ values. (Abstract shortened by UMI.)

Acceleration of the regeneration process is necessary for severe full-thickness skin injuries, such as burns, trauma or chronic ulcers. The aim of this dissertation was to investigate the influence of bone marrow-derived stem cells on skin regeneration using full-thickness wound model in BALB/c mouse. Lin¯ cells were isolated and identified. Regenerative properties of purified cells were evaluated in vitro. Wound healing after Lin¯ cells transplantation was examined histologically and cytokine gene expression was determined by quantitative RT-PCR. The results revealed that Lin¯ population is heterogeneous and contains distinct undifferentiated cell types. In vitro experiments showed that skin tissue extracellular matrix components enhance proliferation and migration of Lin¯ cells. Histologic analysis indicated that inhibition of inflammation, re-epithelization, formation of skin appendages and extracellular collagen, were most effective after cell transplantation with type I collagen. Quantitative analysis revealed that transplanted Lin¯ cells decrease expression of proinflammatory cytokine TNF-α and increase anti-inflammatory IL-10 expression. Also, gene expression of growth factors TGF-β and VEGF, which enhance regeneration, was improved in different healing phases. The results obtained provide additional knowledge for possible therapeutic application of stem/progenitor cells in difficult-to-heal wounds. / Odos žaizdų regeneracijos skatinimas yra aktualus pažeidus didelį paviršiaus plotą bei giliuosius audinio sluoksnius. Šio darbo tikslas buvo ištirti kaulų čiulpų Lin¯ ląstelių įtaką odos regeneracijai in vivo, taikant BALB/c linijos pelių visų odos sluoksnių pažeidimo modelį. Darbo metu buvo išskirtos bei identifikuotos Lin¯ ląstelės. Jų regeneracinės savybės buvo įvertintos in vitro. Buvo atlikta histologinė gyjančio audinio analizė bei nustatyti citokinų genų raiškos pokyčiai. Rezultatai parodė, kad Lin¯ ląstelių populiacija yra heterogeniška, sudaryta iš nediferencijuotų ląstelių. Nustatyta, kad odos tarpląstelinės medžiagos komponentai skatina Lin¯ ląstelių proliferaciją ir migraciją in vitro. Histologinė analizė parodė, kad po ląstelių transplantacijos I tipo kolageno gelyje anksčiausiai prasideda uždegimo slopinimas, greičiausiai baigiamas reepitelizacijos procesas, atsinaujinusioje dermoje susiformuoja tvarkingai išsidėsčiusios pagalbinės odos struktūros bei natyviam audiniui būdingas tarpląstelinės medžiagos kolagenas. Atlikus kiekybinę analizę nustatyta, kad po Lin¯ ląstelių transplantacijos sumažėja uždegiminio citokino TNF-α raiška bei padidėja priešuždegiminio IL-10 raiška. Skirtingose gijimo stadijose padidėja augimo veiksnių, skatinančių audinio regeneraciją, TGF-β ir VEGF genų raiška. Gauti rezultatai papildo žinias, reikalingas vystyti metodus, skirtus sutrikusio gijimo žaizdų regeneracijos skatinimui.

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Background. Recently rodent and porcine bone marrow stromal cells (MSCs) have been reported to be uniquely immune tolerant. In order to confirm these findings in human cells, we tested the hypothesis that human MSCs are also immune tolerant, such that they can be useful as "universal donor cells" for myocardial regenerative therapy. / Methods. Immunocompetent female rats underwent left coronary ligations (n=90). They were randomized into 3 groups. In Group I, lac-Z labeled male human MSCs were implanted into the peri-infarcted area. In Group II and III isogenic rat MSCs or culture medium were injected respectively. Echocardiography was carried out to assess cardiac function, and the specimens were examined serially for up to 8 weeks with immunohistochemistry, FISH and PCR to examine MSCs survival and differentiation. / Results. Human MSCs were found to survive within the rat myocardium without immunosuppression. This was confirmed by PCR and FISH test. No cellular infiltration characteristic of immune rejection was noted. Some of these cells appeared to express cardiomyocyte-specific markers such as troponin-Ic and connexin-43. Furthermore, the implanted MSCs significantly contributed to the improvement in ventricular function and attenuated LV remodeling. / Conclusions. Human MSC survived within this xenogeneic environment, and contributed to the improvement in cardiac function. Our findings support the feasibility of using these cells as "universal donor cells" for xeno- or allo-geneic cell therapy, as they can be tested, prepared and stored well in advance for urgent use. Allogeneic MSCs from healthy donors may be particularly useful for severely ill or elderly patients whose own MSCs could be dysfunctional. / Plusieurs études ont récemment démontré la tolérance immunologiquedes cellules souches stromales (CSS) issues de rongeurs et de porcinés. Pour confirmer cesrésultats chez les cellules humaines, l'étude actuelle évalue l'effet des CSS humaines sur larégénération du myocarde chez des rats immunocompétents et étudie la possibilité d'utiliserces CSS comme « donatrices universelles» à la suite d'un infarctus.