Autologous Hematopoietic Stem Cell Transplantation for Adults With Acute Myeloid Leukemia.

BACKGROUND: Hematopoietic stem cell transplantation (HSCT) remains the most efficacious therapy in patients with acute leukemia. For older patients and those lacking a related HLA-compatible donor, autologous transplantation (auto-HSCT) is a valid alternative therapeutic option. METHODS: From 1997 until 2014 in the Department of Hematooncology and Bone Marrow Transplantation, Medical University of Lublin, Poland, 29 auto-HSCT were performed in patients with acute myeloid leukemia (AML; 15 men and 14 women; median age, 52.2 years). The following FAB types of AML were diagnosed: M0, 3; M1, 4; M2, 6; M4, 10; and M5, 6. Patients with AML were classified into 3 cytogenetic prognostic groups: high risk, 9; intermediate risk, 16; and low risk, 4. Twenty-five were in first complete remission and 4 in second complete remission. The peripheral HSCs mobilized after chemotherapy (mainly second course of consolidation) and granulocyte colony-stimulating factor were the source of the stem cells in all cases. The median number of infused CD34+ cells was 3.58 × 10(6)/kg. The conditioning regimen was busulfan and cyclophosphamide in all patients with AML. The intravenous form of busulfan was applied in the last 15 patients. RESULTS: The median time for absolute neutrophil count recovery >0.5 × 10(9)/L and for platelet count >20.0 × 10(9)/L was 12 and 16.5 days, respectively. Treatment-related mortality rate in the whole group was 3.4% (1 patient with sepsis in the aplastic period). The median follow-up time of survivors was 21.9 months (range, 11.7-142.4). The 3-year projected disease-free survival and overall survival rates were 60% and 68%, respectively. CONCLUSIONS: Our data confirm that auto-HSCT is a valuable therapeutic option for patients with AML, especially older patients and those lacking related HLA-compatible donors.

Isotopic composition and identification of the origins of individuals buried in a Neolithic collective grave at Bronocice (southern Poland).

The oxygen present in a human organism comes from numerous sources, but the major factor that causes variation in the isotopic composition of this element in a tissue is available drinking water. The isotopic ratio of oxygen in an organism's tissue, including that found in bones and teeth, reflects the isotopic oxygen composition typical for the area where a given individual developed and lived. Of particular interest with regard to this issue were a series of skeletons from the multiple grave discovered at the Funnel Beaker-Baden settlement at Bronocice (southern Poland). The question therefore arose whether the specimens buried in this grave were part of the local community. The oxygen isotope level was established using apatite isolated from bones or teeth. A femur and root dentine samples taken from permanent teeth were subjected to oxygen isotope analysis. The oxygen isotope level of the site was established on the basis of local water precipitation and measurements taken from the oxygen isotope concentration in apatite samples isolated from the bones of animals co-occurring with the studied human group. It has been found that the oxygen isotope levels in the bones and dentine of almost all the analysed specimens from the excavated site at Bronocice were within the established range for the area's environment, providing evidence for their local origin. Thus, it can be assumed that the analysed group inhabiting the macrosettlement at Bronocice during the Funnel Beaker phase of the Baden culture was most probably of local origin.

Trace element analysis of Bronze Age skeletal and crematory graves from southern Poland for diet reconstruction.

The basic aim of the present study was to determine the diet of human populations dating from the Bronze Age and the time of Roman influence. The osseous material under examination came from skeletal and crematory burials of the Lusatian and Przeworks Cultures, found in a cemetery at Opatów, Klobuck District, Silesian Province. Three elements: strontium, zinc and calcium were chosen as basic diet determinants. The three elements and the proportions between them are the most frequently used factors that permit a description of the relative proportion of animal and plant protein in a diet. It was assumed that the differentiation of burial ritual was paired with a diverse mode and quality of nourishment. Interdisciplinary osteological analyses, based on physicochemical studies of the odontologic material in the context of archaeological data (culture affiliation, type of burial, grave furnishings), permit a complex analysis of the issues connected with the biology of human groups, their demographic structure or, eventually, paleostratigraphy of primeval communities. It has been found that the type of burial and the richness of grave furnishings are most closely connected with the ultimate differentiation of Zn and Ca concentrations and the value of the Sr/Zn ratio. Because the richly furnished graves are at the same time mostly skeletal burials, it cannot be unequivocally stated which of the above-mentioned factors is of paramount importance. It has also been demonstrated that representatives of the Przeworsk Culture, chronologically younger than the Lusatian one that inhabited the same geographic region, show a lower Zn concentration and a higher Sr/Zn ratio.

Phenotypic changes of monocytes induced by HIV-1 gp120 molecule and its fragments.

Several phenotypic and functional changes of monocytes (M phi) have been described in HIV-1+ subjects and AIDS patients. Some of these changes that are pertinent for immunopathogenesis of the disease may be induced by HIV-1 envelope glycoprotein 120 (gp120). In the present study the effect of recombinant full length gp120 (FLgp120) and its two fragments: rp120cd (aa 410-511) and rp120 (aa 446-511) on the expression of the surface molecules of M phi cultured in vitro was determined. The FLgp120 and rp120cd caused upregulation of CD14 and CD44. The rp120cd peptide significantly increased the expression of CD16 (Fc gamma receptor type III) and TNF receptor type II. In contrast, the rp120 downregulated HLA-DR, CD64 (Fc gamma RI), interferon gamma receptor and induced IL-10 production by M phi. This study indicates that gp120 molecule and its fragments may induce several phenotypic changes of M phi in particular the increased proportion of CD14+CD16+ cells that is observed in the blood of AIDS patients. These results provide further evidence for variable response of M phi to gp120 which may explain the variability of phenotypic changes and heterogeneity of M phi subsets seen in HIV-1 disease.

Interaction of HIV-1 gp120 molecule fragments with human monocytes: different requirements for tumor necrosis factor-alpha and IL-6 production.

The HIV-1 gp120 recombinant protein fragment encompassing aa residues 410-511, that contains the CD4 binding region (rp120cd), and fragment aa 446-511, which lacks the sequence responsible for CD4 binding (rp120), were synthesized to study their ability to induce TNF synthesis in human monocytes. The rp120cd stimulated TNF alpha secretion by monocytes while the rp120 and full-length recombinant protein (FL gp120), used as control, failed to do so. However, FL gp120 stimulated peripheral blood mononuclear cells (PBMC) and lymphocytes for TNF production and this was inhibited by anti-CD4 MAb. The rp120cd also caused TNF secretion by PBMC that was not blocked by this antibody. Furthermore, FL gp120 but not rp120cd inhibited anti-CD4 mAb binding to CEM cells. Hence, FL gp120 may cause TNF release from lymphocytes by binding to CD4, while rp120cd interacts with monocytes but not lymphocytes and induces TNF production by a mechanism not involving CD4 binding. Unexpectedly, FL gp120 but not rp120cd stimulated IL-6 secretion and IL-6 mRNA synthesis in monocytes. The FL gp120-induced production of IL-6 by monocytes was inhibited by anti-CD4 monoclonal antibody (MAb). Thus, there may be different requirements for TNF induction in lymphocytes and monocytes stimulated with various preparations of gp120 and for the selective induction of cytokines in monocytes. The enhanced production of TNF in HIV infection and AIDS may involve distinct cellular sources and different mechanisms.

Modulation of antigen-presenting capacity of human monocytes by HIV-1 GP120 molecule fragments.

The two fragments of HIV-1 gp120 molecule were synthesized to study their interaction with human monocytes. Previous observations indicated that recombinant gp120 fragment (aa residues 410-511) encompassing CD4 binding region (rp120cd) induced tumour necrosis factor alpha (TNF) production in monocytes, while a similar fragment (rp120) not containing the CD4 binding sequence (aa 446-511) was inactive. This paper shows that rp120cd depressed monocyte ability to present antigen (PPD) to autologous T lymphocytes while rp120 was noninhibitory. The rp120cd interacted with monocytes but not T lymphocytes. Anti-TNF receptor type A antibody (utr-1) prevented the depression of antigen presentation caused by rp120cd, which suggested a role for TNF and its receptor. The depression of antigen presentation was seen only when monocytes were treated with rp120cd before, but not after, pulse with antigen. Parallel changes were observed in PPD-induced IL-6 production. Thus, induction of TNF by gp120 may be associated with impairment of antigen-presenting capacity of monocytes seen in AIDS patients.

Synthesis of a gene encoding bovine substance P precursors and its expression in Escherichia coli.

Using an efficient Escherichia coli expression system, we have been able to obtain the precursor of substance P, alpha-preprotachykinin (alpha PPT). The alpha PPT protein is produced in E. coli as a fusion to beta-galactosidase, and accumulates in the cytoplasm as insoluble inclusion bodies. We also produced protachykinin (alpha PT), i.e., alpha PPT without a signal peptide. Further purification and characterization of the alpha PPT and alpha PT polypeptides strongly suggest that fully purified products can be obtained using our procedures.

Modified osmotic fragility test for the laboratory diagnosis of hereditary spherocytosis.

A modified osmotic fragility test, based on measurement of hemolysis in four hypotonic NaCl solutions and logarithmic linearization of osmotic fragility curve is, like the "Pink test," a specific and sensitive test for the laboratory diagnosis of hereditary spherocytosis.

Modified end-point glycerol hemolysis assay as a screening test for hereditary spherocytosis that requires no venipuncture.

The high specificity of the "pink test" for the detection of hereditary spherocytosis has been confirmed. A modification of the test is proposed that requires only 10 microliter of blood taken without anticoagulant (a "direct pink test"), thus eliminating the necessity of venipuncture, especially cumbersome in newborns and infants.