Autologous and Allogeneic Skin Cell Grafts in the Treatment of Severely Burned Patients: Retrospective Clinical Study.

BACKGROUND: Transplantation of skin cells (keratinocytes and fibroblasts) cultured in vitro is a method of choice for the treatment of severe and extensive burns in patients with a deficit of donor sites for free split-thickness skin grafts, and when the grave medical condition of the patient excludes the possibility of an operation under general anesthetic. Appropriate amounts of keratinocytes and/or fibroblasts cultured in vitro are grafted as a suspension in platelet-leukocyte-rich gel directly on the prepared acceptor site. Approximately 3 weeks are needed for autologous cell culture to grow. Allogeneic cells are obtained from patients who died before their own autologous cell transplantation. Therefore allogeneic cells are considered as ready to use product. The aim of the study was to evaluate the efficiency of in vitro cultured autologous/allogeneic skin cell grafts in the treatment of burns. MATERIALS AND METHODS: In this study a group of 68 patients hospitalized in the Dr Stanislaw Sakiel Centre for Burn Treatment in Siemianowice Slaskie and treated with in vitro cultured skin cells suspended in platelet-leukocyte-rich gel were analyzed. RESULTS: Autologous/allogeneic keratinocytes and fibroblasts transplantation hastens wound closure. CONCLUSION: A major factor in burn treatment is early application of skin cells, so allogeneic cells are more appropriate, because these cells are an on-shelf product. It is especially important in groups of patients with third-degree burn greater than 40%. Allogeneic cells application does not increase hospitalization length in comparison to autologous cells, meaning usage of allogeneic cells in burns treatment is as efficient as autologous cells.

Experience in Using Fetal Membranes: The Present and New Perspectives.

INTRODUCTION: The placenta is an accessible source of tissues for transplantation. Placental transplants have been used in wound treatment because of the basic function of the placenta and its nutritious properties and structure. PATIENTS AND METHODS: The aim of this work is to present the clinical usage of fetal membranes, including human amnion, on the basis of the burn treatment center's experience. The clinical use of amnion and different types of placental transplants are described. The initial results of research work within the MEDPIG project are presented regarding the application of placenta from transgenic pigs as a source of tissues for transplantation. RESULTS: From August 2011 to March 2017, 252,592 cm2 of biostatic human amnion transplants were prepared in our tissue bank. During this period they were transplanted to 528 patients, including 10 patients with Lyell syndrome. Initial studies were conducted in which placentas were collected from 5 transgenic pigs and 27,426 cm2 of amniotic grafts were prepared from them. DISCUSSION: The authors' own experience as well as the literature confirm the extraordinary efficiency of transplants prepared from placental tissues, especially from the amniotic membrane. CONCLUSIONS: The clinical effects confirm the effectiveness of using human amnion in wound treatment. Amniotic transplant is a new treatment standard in toxic epidermal necrolysis TEN (Lyell's syndrome), which has found confirmation in very good clinical outcomes. The collected placentas from transgenic animals enabled the preparation of significantly more grafts than in the case of human material, which is a great advantage of this source of placenta over human tissues.

Infections in the tissue material and their impact on the loss of transplants in the Laboratory of in vitro Cell and Tissue Culture with Tissue Bank in the years 2011-2015.

Radiation sterilization eliminates microbiological infections but causes the degradation of the cell factor. The negative result of microbiological examination for tissue transplants is one of the conditions for approval for distribution in patients. The study attempts to verify impact of the presence of microbes onto material for transplant loss. In the 2011-2015 period, we analyzed 293 donors of skin and amnion. Microbiological sampling was performed. The total of 21 strains of bacteria, molds and fungi was identified in collected tissue. The widest spectrum of strains was found in skin (17), followed by amnia (8). The total number of positive findings was 147 and was again highest in skin (129), while the number of positive findings in amnia was 18 only. The general percentage of fungal infections was very low. The presence of fungal strains was only observed in allogeneic skin (2%). Large number of microorganisms isolated from the skin before sterilization was observed, so it seems impossible to use allogeneic intravital skin. However, the intravital application of allogeneic amnion obtained from cesarean section remains to be considered.

Thermoresponsive poly[tri(ethylene glycol) monoethyl ether methacrylate]-peptide surfaces obtained by radiation grafting-synthesis and characterisation.

This report demonstrates the feasibility of radiation grafting for the preparation of polymer layers functionalised with short peptide ligands which promote cell adhesion. Thermoresponsive poly [tri(ethylene glycol) monoethyl ether methacrylate] (PTEGMA) layers were synthesised on a polypropylene substrate by post-irradiation grafting. A cell adhesion moiety, the CF-IKVAVK peptide modified with a methacrylamide function and a fluorescent label were introduced to the surface during the polymerisation process. The amount of CF-IKVAVK was easily controlled by changing its concentration in the reaction mixture. The changes in the surface composition, morphology, philicity and thickness at each step of the polypropylene functionalisation confirmed that the surface modification procedures were successful. The increase in environmental temperature above the cloud point temperature of PTEGMA caused a decrease in surface philicity. The obtained PTEGMA and PTEGMA-peptide surfaces above TCP were tested as scaffolds for fibroblast sheet culture and temperature induced detachment.

A model to simulate pumping tests in wells with high loss and backflow.

Traditional methods of analyzing pumping tests in single wells fail when the well loss is very high due to a low transmissivity skin. Because of the restricted rate at which water can enter a high loss well from the aquifer, well casing storage becomes a significant factor. Additionally, if a slug of water enters the well from the pump column immediately after the pump is switched off, it has a long-lasting significant effect on the recovering water level in the well because it cannot be absorbed rapidly by the aquifer. A theoretical model is derived here that simulates the water level in a well in these circumstances. In the model, the continuously changing rate of water inflow from the aquifer to the well is approximated by a step function with a finite difference time step. It is demonstrated by a real example that the model can be applied easily to analyze pumping tests, including tests with a varying pumping rate. The analysis confirms suspected high well loss, calculates the unknown rate of backflow, and determines the aquifer's transmissivity.

Unconfined linear flow to a horizontal well.

The validity of a previously proposed but untested modification to equations for flow to a horizontal well is assessed using a specially developed finite-difference model. This modification extends confined flow equations to allow the head in the well and the saturated depth at the well to be estimated in unconfined conditions. The study is limited to the case of two-dimensional flow with no flow in the direction parallel to the line of the well. The results show that the modified equations for both a finite unconfined aquifer and, by inference, an infinite unconfined aquifer are adequately accurate for practical application.

Comparison of continuous and cyclic pumping from a well.

Two practically applicable pumping patterns for wells are: (1) continuous pumping at a sustainable constant rate; and (2) pumping at a sustainable constant rate for a fixed period every day (cyclic pumping). Theoretical analysis shows that with the cyclic pumping pattern the pump can be run at a higher rate than with continuous pumping. However continuous pumping at the maximum continuously sustainable constant rate produces considerably more water. If the objective is to produce as much water as possible, it is important to use a pump designed for running at the maximum continuously sustainable rate.

Selective inhibition of prolactin gene transcription by the ETS-2 repressor factor.

Regulation of prolactin gene transcription requires cooperative interactions between the pituitary-specific POU domain protein Pit-1 and members of the ETS transcription factor family. We demonstrate here that the ETS-2 repressor factor (ERF) is expressed in pituitary tumor cells and that overexpression of recombinant ERF inhibits prolactin promoter activity, but not the closely related growth hormone promoter. In non-pituitary cell lines, coexpression of ERF disrupts the cooperative interactions between Pit-1 and ETS-1 and blocks the induction of Pit-1-dependent prolactin promoter activity by cAMP. The potential role of ERF in the inhibitory response of the prolactin promoter to dopamine was examined using pituitary tumor cells stably expressing dopamine D2 receptors. The inhibitory responses of the prolactin promoter to ERF and dopamine are additive, suggesting that ERF has a complementary role in this hormonal response. A single Pit-1 DNA-binding element from the prolactin promoter is sufficient to reconstitute the inhibitory response to ERF. DNA binding analysis using either a composite Pit-1/ETS protein-binding site or a Pit-1 element with no known affinity for ETS proteins revealed that ERF interferes with Pit-1 binding. Together, these results demonstrate that ERF is a specific inhibitor of basal and hormone-regulated transcription of the prolactin gene and suggest a new level of complexity for the interaction of ETS factors with Pit-1 target genes.

Dual-function reporter protein for analysis of gene expression in living cells.

The firefly luciferase (Luc) protein and the jellyfish green fluorescent protein (GFP) are two commonly used molecular reporters that can be detected noninvasively in living cells. The properties that make GFP or Luc useful for a particular experimental application are quite distinct. A recombinant protein with both fluorescent and bioluminescent characteristics might take advantage of the strengths of both reporters. An expression vector encoding a chimeric protein in which GFP was tethered to Luc through a 19-amino acid linker was prepared and characterized. Western blotting with antibodies specific for either GFP or Luc showed that a protein of appropriate size was expressed in transfected cells. Fluorescence microscopy revealed bright green fluorescence from transfected cells, indicating proper formation of the GFP chromophore. Luc enzymatic activity in protein extracts from transfected cells showed that Luc was fully functional. The treatment of living cell cultures stably expressing the GFP-Luc fusion protein with the protein translation-inhibitor cycloheximide (Chx) was used to show that the half-life for Luc protein activity was approximately 2 h at 37 degrees C. The utility of this dual-function reporter protein was shown by the identification of single living cells expressing the chimeric protein within a population by fluorescence microscopy, followed by quantification of Luc activity from the same living cells.

[Rectal cancer in clinical material. Effect of advanced clinical staging, tumor location and type of surgical treatment on long term results]. / Rak odbytnicy w materiale kliniki. Wplyw stopnia zaawansowania klinicznego, umiejscowienia guza i sposobu leczenia chirurgicznego na odlegle wyniki.

The aim of the research is presentation of patients' material, operated on in The Clinic in the years 1965-1993 because of rectal cancer. The material makes 1363 patients, where group of 214 patients was separated from, treated in the years 1989-1993, basing on modern system of medical examinations qualifying for the surgical intervention and included by intensive prospecting monitoring. At investigative periods 1965-1989 and 1989-1993 the transition of rectal Dukes' stages on A and B interest was noticed, this admitted to increase the whole number of local excision et anterior rectum resection and improve distant treatment results, five years' survival measured. Percentage of patients with inoperable clinical staging, at periods: 1965-1989 and 1989-1993, is similar, in spite of considerable development of diagnostic methods.

[Inflammatory tumors of the colon--diagnostic and therapeutic problem in clinical materials]. / Guzy zapalne jelita grubego--problem diagnostyczny i terapeutyczny w materiale kliniki.

38 patients with inflammatory tumours of the large bowel (ITLB) were studied, it is 2.4% of all patients operated in our department for the large bowel tumours. 28 patients (73.7%) underwent elective surgery, 10 (26.3%) had emergency operations. One-stage operations were performed in 26 cases (68.4%) and two-stages operations in 12 (31.6%). Permanent colostomies were performed in 4 patients (10.5%). Histological examination indicated chronic colitis in 19 cases (50.0%), mostly on divercular disease basis, Crohn's disease in 7 (18.4%), profound ulcerative colitis in 5 (13.1%), actinomycosis in 3 (7.9%) and typhlitis in 2 (5.3%).

[Abdominal resection of rectal cancer]. / Wyciecie brzuszne odbytnicy z powodu raka.

Preservation of anal sphincters in the surgical treatment of rectal cancer is often controversial as far as an extent of surgery, incidence of complications and an increase in mortality rate in perioperative period are concerned. The study was aimed at results of the retrospective analysis of patients, who undergone abdominal excision of the rectum for malignant tumours. Totally 136 patients were operated. Perioperative mortality rate was 5.14%. Totally 26.4% of patients survived for at least 5 years, including 80% of patients operated in A stage of cancer, according to Duke, and 43.3% of patients with tumour in B stage. An emphasis is on the necessity to leave a 2-centimeter margin of healthy tissues around the tumour, provided, that the result of intraoperative histological examination was negative. Such a margin is sufficient for anastomosis. Low anastomoses do not produce early complications and recurrences.