Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 36
Filter
Add more filters










Publication year range
1.
Spectrochim Acta A Mol Biomol Spectrosc ; 314: 124177, 2024 Jun 05.
Article in English | MEDLINE | ID: mdl-38554690

ABSTRACT

The SERS response of adenine is one of the most studied, due to its outstanding exaltation. However, the spectra obtained strongly differ according to the experimental conditions and still remain not well understood. To be able to search for the presence of this molecule in complex environments, it is essential to better understand the SERS response of adenine alone. After a brief presentation of the literature on the subject, we present results suggesting that the experimental spectra would result from the overlap of different spectroscopic signatures, that may probably be due to different non-covalent interactions or different electromagnetic fields experienced by adenine molecules. An independent component analysis is reported. Our results underline the difficulty to precisely analyze the experimental data, the need to continue this research and to constitute data banks that would allow comparing the spectra obtained in different laboratories according to the experimental conditions.

2.
Geobiology ; 16(1): 49-61, 2018 01.
Article in English | MEDLINE | ID: mdl-29076282

ABSTRACT

Cyanobacteria have long been thought to induce the formation of Ca-carbonates as secondary by-products of their metabolic activity, by shifting the chemical composition of their extracellular environment to conditions favoring mineral precipitation. Some cyanobacterial species forming Ca-carbonates intracellularly were recently discovered. However, the environmental conditions under which this intracellular biomineralization process can occur and the impact of cyanobacterial species forming Ca-carbonates intracellularly on extracellular carbonatogenesis are not known. Here, we show that these cyanobacteria can form Ca-carbonates intracellularly while growing in extracellular solutions undersaturated with respect to all Ca-carbonate phases, that is, conditions thermodynamically unfavorable to mineral precipitation. This shows that intracellular Ca-carbonate biomineralization is an active process; that is, it costs energy provided by the cells. The cost of energy may be due to the active accumulation of Ca intracellularly. Moreover, unlike cyanobacterial strains that have been usually considered before by studies on Ca-carbonate biomineralization, cyanobacteria forming intracellular carbonates may slow down or hamper extracellular carbonatogenesis, by decreasing the saturation index of their extracellular solution following the buffering of the concentration of extracellular calcium to low levels.


Subject(s)
Calcium Carbonate/metabolism , Cyanothece/metabolism , Calcium/metabolism , Culture Techniques , Cyanothece/growth & development
3.
Leukemia ; 29(5): 1084-91, 2015 May.
Article in English | MEDLINE | ID: mdl-25627637

ABSTRACT

In 2008, a European registry of relapsed acute promyelocytic leukemia was established by the European LeukemiaNet. Outcome data were available for 155 patients treated with arsenic trioxide in first relapse. In hematological relapse (n=104), 91% of the patients entered complete hematological remission (CR), 7% had induction death and 2% resistance, 27% developed differentiation syndrome and 39% leukocytosis, whereas no death or side effects occurred in patients treated in molecular relapse (n=40). The rate of molecular (m)CR was 74% in hematological and 62% in molecular relapse (P=0.3). All patients with extramedullary relapse (n=11) entered clinical and mCR. After 3.2 years median follow-up, the 3-year overall survival (OS) and cumulative incidence of second relapse were 68% and 41% in hematological relapse, 66% and 48% in molecular relapse and 90 and 11% in extramedullary relapse, respectively. After allogeneic or autologous transplantation in second CR (n=93), the 3-year OS was 80% compared with 59% without transplantation (n=55) (P=0.03). Multivariable analysis demonstrated the favorable prognostic impact of first remission duration ⩾1.5 years, achievement of mCR and allogeneic or autologous transplantation on OS of patients alive after induction (P=0.03, P=0.01, P=0.01) and on leukemia-free survival (P=0.006, P<0.0001, P=0.003), respectively.


Subject(s)
Antineoplastic Agents/therapeutic use , Arsenicals/therapeutic use , Leukemia, Promyelocytic, Acute/drug therapy , Oxides/therapeutic use , Adolescent , Adult , Aged , Aged, 80 and over , Arsenic Trioxide , Cell Differentiation , Child , Child, Preschool , Disease-Free Survival , Europe , Female , Humans , International Cooperation , Leukemia, Promyelocytic, Acute/pathology , Male , Middle Aged , Prospective Studies , Recurrence , Registries , Treatment Outcome , Young Adult
5.
Rev Med Suisse ; 9(404): 1998-2001, 2013 Oct 30.
Article in French | MEDLINE | ID: mdl-24313051

ABSTRACT

Performing a complete blood count analysis is a daily routine necessary for a good care of patients. Nowadays, modern blood analyzers provide on top of classical blood values, several additional parameters. In this paper, using short case presentations, we discuss how to interpret these results and integrate them in the clinical context.


Subject(s)
Blood Cell Count/methods , Patient Care/methods , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Practice Patterns, Physicians'
6.
Appl Psychophysiol Biofeedback ; 34(2): 105-11, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19308725

ABSTRACT

In this paper we examined the influence of repetition of weight-related sentences on the involuntary pressure forces of the forearms, when in a relaxed state. These forces were involuntary oscillations, exerted by muscle movements of the breathing-cycle and muscle movements of the arm on force sensors. We constructed a linguistic bio-mechanical system (Ablasmi), where in each arm of a padded relaxation chair contained special sensors that were specifically designed to detect the pressure forces of each forearm while participants listened to the specific recorded weight-related sentences. In this experiment we used some classic sentences, such as "your right/left arm is heavy," inspired by Autogenic Training (Schultz in Le training autogene. Paris, PUF, 1974) and some sentences, such as "your right/left arm is made of lead," inspired by Grossmann's relaxation technique. We observed that when the recorded sentences were directed to the right arm there was a significant increase of involuntary pressure forces on the right forearm. Additionally, we observed the same effect on the left forearm for the sentences directed to the left forearm. Thus, we believe we have provided experimental evidence of a definite linguistic effect of weight-related sentences on the involuntary pressure forces of the forearms. The implications of these findings are discussed.


Subject(s)
Forearm/physiology , Language , Relaxation Therapy , Relaxation/physiology , Adult , Arm/physiology , Dancing/physiology , Electromyography , Female , Functional Laterality/physiology , Humans , Male , Pressure , Respiratory Mechanics , Young Adult
7.
Langmuir ; 23(5): 2463-71, 2007 Feb 27.
Article in English | MEDLINE | ID: mdl-17274633

ABSTRACT

The adsorption of proteins is the first process to take place when a solid is immersed in a biological fluid; though not yet thoroughly understood at a molecular level, this process is also known to be strongly influenced by the presence of salt in solution or by pH changes. In the present work, poly-L-glutamic acid (PG) and poly-L-lysine (PL) were selected to mimic the behavior of some protein fragments. Their adsorption was investigated by infrared spectroscopy in various modes, both on planar and on divided (powder) surfaces of aluminum oxide. These two peptides were shown to have different behaviors when adsorbed from solutions with or without CaCl2 and at various pH values. Polarization modulation-reflection absorption infrared spectroscopy, applied in a special cell designed to characterize the solid surface in contact with the liquid, enabled the observation of the influence of pH and salts upon polypeptide adsorption. At pH values higher than 5 and in the presence of CaCl2 in solution, a net increase of the PG adsorbed amount is observed, whereas no such effect could be detected for PL. Specific interactions between the COO- groups on the side chains and the surface, or between those of two different molecules, was inferred. Interestingly, similar conclusions could be drawn for the surface of alumina powders contacted with solutions of PG and PL and characterized by attenuated total reflectance IR. This work demonstrates the potential for IR investigations of solid oxide-liquid interfaces combining the study of planar and finely divided surfaces.


Subject(s)
Aluminum Oxide/chemistry , Calcium Chloride/chemistry , Peptides/chemistry , Proteins/chemistry , Spectrophotometry, Infrared/methods , Adsorption , Chemistry, Physical/methods , Hydrogen-Ion Concentration , Models, Chemical , Nanostructures , Polyglutamic Acid/chemistry , Powders , Spectroscopy, Fourier Transform Infrared , Surface Properties
8.
J Thromb Haemost ; 5(4): 826-34, 2007 Apr.
Article in English | MEDLINE | ID: mdl-17229052

ABSTRACT

BACKGROUND: Blood-derived endothelial progenitor cells (EPC) have been used to treat ischemic disease. However, the number of EPC that can be obtained from adult blood is limited. OBJECTIVE: To characterize endothelial-like cells obtained from human bone marrow and determine their ability to stimulate new blood vessel formation in vivo. METHODS: Mononuclear cells (MNC) were isolated from human bone marrow or umbilical cord blood and cultured in endothelial growth medium (EGM-2). Mesenchymal stem cells (MSC) were isolated from bone marrow and induced to differentiate into endothelial-like cells (MSCE), or adipocytes or osteocytes by growth in EGM-2, adipogenic or osteogenic medium. RESULTS: Cells obtained by culturing bone marrow MNC in EGM-2 formed cord- or tube-like structures when grown on Matrigel(TM) and expressed several endothelial marker proteins. However, cell morphology and the profile of endothelial marker protein expression were different from those of cord blood-derived EPC (cbEPC). Cells with a similar phenotype were obtained by differentiation of MSC into MSCE, which was accompanied by an increase of endothelial marker proteins and a diminished capacity to differentiate into adipocytes. Subcutaneous implantation of MSCE in collagen plugs in non-obese diabetic-severe combined immunodeficient (NOD-SCID) mice resulted in formation of functional blood vessels that had incorporated the MSCE. CONCLUSIONS: Our results show that MSCE and cbEPC are different cell types. The formation of functional blood vessels by MSCE, combined with high yields and a reduced capacity to differentiate into other cell types compared with MSC, makes these cells potentially useful for autologous therapy of ischemic disease.


Subject(s)
Endothelium, Vascular/cytology , Mesenchymal Stem Cells/cytology , Adipocytes/cytology , Animals , Bone Marrow Cells/cytology , Cell Culture Techniques/methods , Cell Differentiation , Collagen/pharmacology , Drug Combinations , Humans , Laminin/pharmacology , Leukocytes, Mononuclear/cytology , Mice , Mice, SCID , Osteocytes/cytology , Proteoglycans/pharmacology , Stem Cells , Umbilical Cord/cytology
9.
Rev Med Suisse ; 2(91): 2852-7, 2006 Dec 13.
Article in French | MEDLINE | ID: mdl-17236325

ABSTRACT

Acute chest syndrome is frequent in the homozygous sickle cell disease population. It can evolve to an acute respiratory distress syndrome. Pulmonary artery hypertension or chronic lung sequellae are common. The vasoocclusive phenomenon is due to capillary blockade, followed by an activation of inflammation, and adhesion phenomena further increasing the damage. Decreased blood oxygenation leads to an aggravation of the sickle crisis. Nitric oxide disregulation has been recently highlighted. Diagnosis must be suspected in patients presenting with sickle crisis, fever, low blood oxygenation and lung infiltrates. Early antibiotherapy, adequate oxygenation, blood transfusion and erythrocytapheresis are key points for management. Preventive measures such as iterative transfusion-chelation, hydroxyurea or repetitive erythrocytapheresis are all useful.


Subject(s)
Anemia, Sickle Cell/therapy , Respiratory Distress Syndrome/therapy , Acute Disease , Adult , Anemia, Sickle Cell/physiopathology , Humans , Respiratory Distress Syndrome/physiopathology
10.
Chemphyschem ; 6(6): 1061-70, 2005 Jun 13.
Article in English | MEDLINE | ID: mdl-15945067

ABSTRACT

The adsorption of neutral glycine onto amorphous silica was investigated both theoretically and experimentally. DFT calculations were performed at the BLYP-631++G** level using a cluster approach. Several possible configurations involving the formation of H bonds between glycine and one, two, or three silanol groups (SiOH) were considered. The most favorable bonding of glycine with one silanol group (45 kJ mol(-1)) occurs through the COOH moiety, thus forming a cycle in which the CO group is an H-bond acceptor whereas the acidic OH group is an H-bond donor. With two or three silanol groups, additional H bonds are formed between the amine moiety and the silanol groups, which leads to an increased adsorption energy (70 and 80 kJ mol(-1) for two and three silanol groups, respectively). Calculated nu(CO), delta(HNH), and delta(HCH) values are sensitive to the adsorption mode. A bathochromic shift of nu(CO) as compared to the nu(CO) of free glycine (calculated in the 1755-1790 cm(-1) range) is found for glycine in interaction with silanol(s). The more H bonds are formed between the COOH moiety and silanol groups, the higher the bathochromic shift. For delta(HNH), no shift is found for glycine adsorbed on one and two silanol groups (where the amine is either not bound or an H-bond donor), whereas a bathochromic shift is calculated with three silanols when the amine moiety is an H-bond acceptor. Experimental FTIR spectra performed at room temperature for glycine adsorbed at 160 degrees C on Aerosil amorphous silica exhibit bands at 1371, 1423, 1630, and 1699 cm(-1). The experimental/calculated frequencies have their best correspondence for glycine adsorbed on two silanol groups. It is important to note that the forms giving the best correspondence to experimental frequencies are the most stable ones.


Subject(s)
Glycine/chemistry , Silicon Dioxide/chemistry , Adsorption , Models, Theoretical , Spectroscopy, Fourier Transform Infrared
12.
Leukemia ; 18(3): 575-83, 2004 Mar.
Article in English | MEDLINE | ID: mdl-14749701

ABSTRACT

In unperturbed mice, the marrow cell numbers correlate with the stem cell numbers. High levels of long-term marrow engraftment are obtained with infusion of high levels of marrow cells in untreated mice. To address the issue of stem cell competition vs 'opening space', knowledge of total murine marrow cellularity and distribution of stem and progenitor cells are necessary. We determined these parameters in different mouse strains. Total cellularity in BALB/c mice was 530+/-20 million cells; stable from 8 weeks to 1 year of age. C57BL/6J mice had 466+/-48 million marrow cells. Using these data, theoretical models of infused marrow (40 million cells) replacing or adding to host marrow give chimerism values of 7.5 and 7.0%, respectively; the observed 8-week engraftment of 40 million male BALB/c marrow cells into female hosts (72 mice) gave a value of 6.91+/-0.4%. This indicates that syngeneic engraftment is determined by stem cell competition. Our studies demonstrate that most marrow cells, progenitors and engraftable stem cells are in the spine. There was increased concentration of progenitors in the spine. Total marrow harvest for stem cell purification and other experimental purposes was both mouse and cost efficient with over a four-fold decrease in animal use and a financial saving.


Subject(s)
Bone Marrow Cells/cytology , Hematopoietic Stem Cells/cytology , Animals , Blood Cell Count , Cell Separation/methods , Female , Graft Survival , Hematopoietic Stem Cell Transplantation/methods , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL
13.
Leukemia ; 17(9): 1871-9, 2003 Sep.
Article in English | MEDLINE | ID: mdl-12970789

ABSTRACT

Long-term multilineage allochimerism can be obtained in H2-mismatched B6.SJL to BALB/c transplants with host irradiation of 100 cGy, donor spleen cell pre-exposure and costimulator blockade with anti-CD40 ligand (CD40L) antibody. We evaluated this allochimerism approach in murine marrow transplants with different degrees of major histocompatibility complexe (MHC) mismatching; these include: (1) H2-mismatched transplant H2Kk to H2Kb, (2) full haplo-identical transplant H2Kbd to H2Kbk, (3) a partial haplo-identical transplant H2Kd to H2Kbd and (4) an MHC class II mismatch. Levels of chimerism increased up to 12 weeks and then stayed relatively stable up to 1 year after transplant. At 18 weeks post-transplant, the H2-mismatched, haplo-identical, partial haplo-identical and class II-mismatch transplants evidenced 17.9+/-4.4, 40.7+/-0.9, 25.1+/-4.19 and 33.7+/-3.5% donor chimerism, respectively. Dropping the anti-CD40 antibody treatment and spleen cells or changing the schedule of antibody to one injection, in haplo-identical or full-mismatched transplants resulted in no donor-derived chimerism. On the other hand, these still resulted in minor chimerism in class II-mismatched transplants. Lineage analysis of peripheral blood at 6 and 12 months post-transplant demonstrated a significant shift toward increased chimeric lymphocytes and decreased chimeric granulocytes in the full H2 as compared with haplo-identical or class II transplants. Transplantation with anti-CD40L antibody eliminated both graft-versus-leukemia and graft-versus-host disease (GVHD) and delayed lymphocyte infusion did not rescue animals from fatal leukemia. In conclusion, under the conditions of our tolerization regimen, a haplo transplant gives higher engraftment levels than a full H2 mismatch, and despite lower engraftment levels, a class II-mismatched transplant can be successfully accomplished with only 100 cGy and no CD40L blockade.


Subject(s)
Bone Marrow Transplantation , CD40 Ligand/immunology , Graft vs Leukemia Effect/immunology , H-2 Antigens/immunology , Transplantation Tolerance , Animals , Antibodies, Monoclonal , Cell Transplantation , Dose-Response Relationship, Drug , Flow Cytometry , Genetic Variation , Graft Survival/drug effects , Graft Survival/radiation effects , Immunophenotyping , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Inbred CBA , Mice, Inbred DBA , Spleen/cytology , Transplantation Chimera/immunology , Whole-Body Irradiation
14.
Bone Marrow Transplant ; 32 Suppl 1: S19-22, 2003 Aug.
Article in English | MEDLINE | ID: mdl-12931234

ABSTRACT

The marrow hematopoietic stem cell is currently being redefined as to all aspects of its phenotype and its total differentiation capacity. This redefinition now includes its plasticity as to production of nonhematopoietic and hematopoietic cell types, the determinants of its in vivo engraftment potential and its expression of stem cell functional characteristics.


Subject(s)
Bone Marrow Cells/cytology , Hematopoietic Stem Cells/cytology , Pluripotent Stem Cells/cytology , Animals , Cell Cycle , Cell Differentiation , Hematopoiesis , Humans
15.
Ann N Y Acad Sci ; 996: 209-21, 2003 May.
Article in English | MEDLINE | ID: mdl-12799298

ABSTRACT

On the basis of our studies of the fluctuation of the hematopoietic stem cell phenotype with cell cycle trnsit, we hypothesize that the ability of marrow stem cells to convert to nonhematopoietic cells will also vary at different points in the cell cycle. The new biology of stem cells has an impact on many fields including developmental biology and stem cell biology and the clinical potential is enormous.


Subject(s)
Hematopoietic Stem Cells/cytology , Animals , Cell Cycle , Cell Differentiation , Cell Size , Cytokines/pharmacology , Hematopoietic Stem Cells/drug effects , Mice , Time Factors
16.
Int J Hematol ; 74(1): 79-85, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11530810

ABSTRACT

We evaluated the engraftment and the cell cycle status of marrow cells at various times after 5-fluorouracil (5-FU) administration. 5-FU (150 mg/kg) was given to donor male BALB/c mice at 1, 2, 6, or 12 days prior to marrow harvest. The donor cells were then assessed in host nonmyeloablated female mice. Bone marrow engraftment of marrow treated with 5-FU was evaluated and compared to marrow treated with diluent (phosphate-buffered saline) at 3 and 10 weeks after marrow infusion. Our data show a rapid induction of an engraftment defect 1 day after 5-FU, persistence of this defect through day 6, and a recovery by day 12. Experiments using hydroxyurea (which selectively kills cells in the S phase) to determine the cell cycle status indicated that cells that engrafted in post-5-FU marrow were noncycling at days 1, 2, and 12 but cycling at day 6. Post-5-FU bone marrow was also analyzed in vitro by colony assays and its cycling status determined by 3H-thymidine suicide assay. High-proliferative-potential colony-forming cells (HPP-CFCs) and low-proliferative-potential colony-forming cells (LPP-CFCs) decreased rapidly 1 day after 5-FU, with a nadir observed at day 6 for HPP-CFCs and day 2 for LPP-CFCs. By day 12, LPP-CFCs showed a total recovery, but HPP-CFCs were still defective. Significant numbers of HPP-CFCs were cycling, mostly at days 6 and 8 after 5-FU, whereas LPP-CFCs appeared quiescent except at day 2. These results emphasize the importance of timing if post-5-FU marrow is used for gene therapy or marrow transplantation.


Subject(s)
Bone Marrow Cells/drug effects , Bone Marrow Transplantation , Fluorouracil/administration & dosage , Hematopoietic Stem Cells/drug effects , Animals , Blotting, Southern , Cell Cycle/drug effects , Colony-Forming Units Assay , DNA Replication , Drug Administration Schedule , Female , Fluorouracil/pharmacology , Graft Survival , Hydroxyurea/pharmacology , Male , Mice
17.
Blood ; 98(4): 1246-51, 2001 Aug 15.
Article in English | MEDLINE | ID: mdl-11493477

ABSTRACT

High levels of chimerism in syngeneic BALB/c transplants were reported when hosts were exposed to 1 Gy (100 cGy) whole body irradiation (WBI) and infused with 40 x 10(6) marrow cells. The recovery of host stem cells and alterations of enhanced host engraftability at varying times after 1 Gy WBI have now been evaluated in this study. Male BALB/c marrow (40 x 10(6) cells) was infused into female BALB/c hosts immediately or at 6, 12, and 24 weeks after 1 Gy WBI of host female BALB/c mice; engraftment percentages 8 weeks after cell injection at week 0, 6, 12, or 24 were 68% +/- 12%, 45% +/- 15%, 51% +/- 12%, or 20% +/- 8%, respectively. Eight-week engraftment levels in nonirradiated hosts average 7.7%. Conversely, engraftable stem cells measured at 8 weeks postengraftment in 1 Gy--exposed hosts were reduced to 8.6% +/- 3% of nonirradiated mice at time 0, 35% +/- 12% 6 weeks later, 49% +/- 10% at 3 months, and 21% +/- 7% at 6 months. Engraftment was still increased and stem cell decreased 1 year after 1 Gy. Furthermore, the primary cells transplanted into 1 Gy hosts can be serially transplanted, and the predominant effect of 1 Gy is directly on engrafting stem cells and not through accessory cells. These data show that transplantation in 1 Gy mice may be delayed until recovery of hematopoiesis, suggesting strategies in allogeneic transplantation to avoid the adverse effects of cytokine storm. The incomplete recovery of engraftable stem cells out to 12 months indicates that stem cell expansion, especially in patients previously treated with radiomimetic drugs, may not be feasible. (Blood. 2001;98:1246-1251)


Subject(s)
Bone Marrow Transplantation/methods , Graft Survival/radiation effects , Hematopoiesis/radiation effects , Hematopoietic Stem Cell Transplantation , Whole-Body Irradiation , Animals , Bone Marrow Cells/cytology , Bone Marrow Transplantation/standards , Female , Male , Mice , Mice, Inbred BALB C , Time Factors , Transplantation Chimera , Transplantation, Isogeneic/methods , Transplantation, Isogeneic/standards
18.
Ann N Y Acad Sci ; 938: 54-61; discussion 61-2, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11458526

ABSTRACT

The donor stem cell phenotype and host microenvironment determine the outcome of a stem cell transplant. In a series of transplant studies in syngeneic male to female or congenic Ly5.1/Ly5.2 models in which hosts have received no or minimal irradiation (100 cGy), evidence overwhelmingly supports the concept that syngeneic engraftment is determined by stem cell competition. These approaches can be extended to H-2 mismatched allogeneic mouse combination when antigen pre-exposure and CD40-CD40 ligand antibody blockage are employed. A human trial in patients with resistant neoplasia infusing pheresed blood with 10(8) CD3 cells/kg showed that tumor responses and complete chimerism occur with very low levels of CD34+ cells/kg and that the extent of previous treatment is a critical factor in determining chimerism. A major feature of transplants is the phenotype of the donor stem cell. This phenotype shows dramatic reversible plasticity involving differentiation, adhesion protein expression, and engraftment with cytokine-induced cell-cycle transit. Homing is probably also plastic. Marked fluctuations in engraftment capacity are also seen at different points in marrow circadian rhythm.


Subject(s)
Graft Survival , Hematopoietic Stem Cell Transplantation/methods , Transplantation Conditioning/methods , Adolescent , Adult , Aged , Animals , Antibodies, Monoclonal/pharmacology , Antigens, Ly/immunology , Apoptosis/drug effects , CD40 Antigens/physiology , CD40 Ligand/drug effects , CD40 Ligand/physiology , Cell Lineage , Chimera , Circadian Rhythm , Clinical Trials as Topic , Dose-Response Relationship, Radiation , Female , Fluorouracil/pharmacology , Graft Enhancement, Immunologic/methods , Graft Survival/drug effects , Graft vs Host Disease , H-2 Antigens/immunology , Hematopoietic Stem Cells/cytology , Hematopoietic Stem Cells/drug effects , Hematopoietic Stem Cells/radiation effects , Histocompatibility , Humans , In Situ Hybridization, Fluorescence , Lymphocyte Transfusion , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Middle Aged , Neoplasms/therapy , Phenotype , Radiation Chimera , Spleen/cytology , Thalassemia/therapy , Transplantation Conditioning/adverse effects , Whole-Body Irradiation
19.
Blood Cells Mol Dis ; 27(5): 934-7, 2001.
Article in English | MEDLINE | ID: mdl-11783958

ABSTRACT

Stem cells from a variety of tissues have recently been shown to be capable of differentiating into cells characteristic of a separate tissue, apparently in response to microenvironmental signals. This is hierarchical plasticity. We have shown that both human and murine neurosphere cells with potential for differentiating into neurons, oligodendrocytes, and astrocytes can produce hematopoietic stem cells when engrafted into fetal sheep or murine day 3.5 blastocysts, respectively. We have also demonstrated an alternative form of stem cell plasticity: functional plasticity at different points in cell cycle transit and at different phases of a circadian rhythm. We have shown that long-term engraftment varies reversibly as primitive murine stem cells (lineage-negative rhodamine(low) Hoechst(low)) transit the cell cycle under stimulation by interleukin-3 (IL-3), IL-6, IL-11, and steel factor, with engraftment being defective in late S/early G2. Engraftment also varies markedly with circadian time. Presumptive mechanisms for these phenotypic shifts include alteration in adhesion protein expression with consequent changes in marrow homing. Most recently, we have also demonstrated that stem cell differentiation varies markedly with cell cycle transit. There are other features of the hematopoietic stem cell which suggest that it is a highly plastic cell with the ability to rapidly change its membrane phenotype, while exhibiting extraordinary directed motility. These data suggest that cell cycle and circadian plasticity should be considered additional major features of the hematopoietic stem cell phenotype.


Subject(s)
Hematopoietic Stem Cells/cytology , Stem Cell Transplantation , Animals , Cell Cycle/physiology , Cell Differentiation , Cell Lineage , Circadian Rhythm , Hematopoietic Stem Cells/physiology , Mice
20.
J Neurosci Methods ; 95(2): 127-32, 2000 Feb 15.
Article in English | MEDLINE | ID: mdl-10752483

ABSTRACT

We designed a rapid, simple and accurate PCR method to determine sexual identity of mouse fetuses collected on embryonic day 15. A multiplex PCR amplification was used to detect male-specific sequence (Sry) in DNA extracted from fetal livers through SDS denaturation followed by high salt extraction and precipitation. This extraction method resulted in sufficiently purified DNA in < 1 h and was suitable for PCR. The DNA obtained was amplified using a robot thermal cycler for 33 cycles. The reaction was performed in 50 microl, using two sets of primers specific for Sry gene (chromosome Y) and IL3 gene (chromosome 11). Amplification duration was 1.5 h. The assessment of the results was done by electrophoresis in 3% agarose run at high voltage. The 402 bp band (Sry) obtained identifies the male fetuses and the 544 bp product (IL3) confirms the correct amplification of the template DNA. The entire procedure took < 4 h. The specificity of the method was confirmed by fluorescent in situ hybridization using a specific male probe on cultured male and female neural stem cells. This method allowed the preparation and culture of pure male and female neural stem cells from fetal tissue.


Subject(s)
Polymerase Chain Reaction/methods , Sex Determination Analysis , Animals , Cell Culture Techniques , DNA/isolation & purification , Female , Fetus , In Situ Hybridization , Male , Mice , Mice, Inbred BALB C , Neostriatum/cytology , Stem Cells/cytology , Time Factors
SELECTION OF CITATIONS
SEARCH DETAIL
...