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J Biol Chem. 2002 Jul 12;277(28):25323-8. Epub 2002 May 08.
Targeted disruption of spermidine/spermine N1-acetyltransferase gene in mouse embryonic stem cells. Effects on polyamine homeostasis and sensitivity to polyamine analogues.

Niiranen K, Pietila M, Pirttila TJ, Jarvinen A, Halmekyto M, Korhonen VP, Keinanen TA, Alhonen L, Janne J.

A. I. Virtanen Institute for Molecular Sciences, University of Kuopio, P. O. Box 1627, Finland.

We have generated mouse embryonic stem cells with targeted disruption of spermidine/spermine N(1)-acetyltransferase (SSAT) gene. The targeted cells did not contain any inducible SSAT activity, and the SSAT protein was not present. The SSAT-deficient cells proliferated normally and appeared to maintain otherwise similar polyamine pools as did the wild-type cells, with the possible exception of constantly elevated (about 30%) cellular spermidine. As expected, the mutated cells were significantly more resistant toward the growth-inhibitory action of polyamine analogues, such as N(1),N(11)-diethylnorspermine. However, this resistance was not directly attributable to cellular depletion of the higher polyamines spermidine and spermine, as the analogue depleted the polyamine pools almost equally effectively in both wild-type and SSAT-deficient cells. Tracer experiments with [C(14)]-labeled spermidine revealed that SSAT activity is essential for the back-conversion of spermidine to putrescine as radioactive N(1)-acetylspermidine and putrescine were readily detectable in N(1),N(11)-diethylnorspermine-exposed wild-type cells but not in SSAT-deficient cells. Similar experiments with [C(14)]spermine indicated that the latter polyamine was converted to spermidine in both cell lines and, unexpectedly, more effectively in the targeted cells than in the parental cells. This back-conversion was only partly inhibited by MDL72527, an inhibitor of polyamine oxidase. These results indicated that SSAT does not play a major role in the maintenance of polyamine homeostasis, and the toxicity exerted by polyamine analogues is largely not based on SSAT-induced depletion of the natural polyamines. Moreover, embryonic stem cells appear to operate an SSAT-independent system for the back-conversion of spermine to spermidine.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12000764&dopt=Abstract

Genes Dev. 2002 May 1;16(9):1129-38.
WUSCHEL signaling functions in interregional communication during Arabidopsis ovule development.

Gross-Hardt R, Lenhard M, Laux T.

Institute of Plant Biology, University of Zurich, 8008 Zurich, Switzerland.

Coordinating the behaviors of different cell populations is essential for multicellular development. One important example for this can be found in ovule development in higher plants. Ovules give rise to the gametophyte in the distal nucellus and form protective sporophytic organs from the underlying chalaza. We show that the WUSCHEL (WUS) homeobox gene provides a mechanism to coordinate these events. WUS is expressed in the nucellus and our loss- and gain-of-function analyses show that WUS is not only necessary but also sufficient for integument formation from the chalaza. WUS protein is retained in the nucellus, indicating that WUS activity in the nucellus generates a downstream signal that non-cell-autonomously regulates integument initiation in the chalaza. This signal appears to act locally, thus determining the position of organ formation from chalazal cells adjacent to the nucellus. Analysis of WUS and AINTEGUMENTA functions indicates that integument initiation requires inputs from different ovule regions. Together with previous findings for shoot and floral meristems, where WUS signaling establishes a stem cell niche, our results indicate that WUS defines a signaling mechanism that is used repeatedly during plant development in coordinating the behavior of adjacent cell groups.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12000795&dopt=Abstract

Clin Lymphoma. 2002 Sep;3(2):111-6.
Mitoxantrone/ifosfamide/etoposide salvage regimen with rituximab for in vivo purging in patients with relapsed lymphoma.

Emmanouilides C, Lill M, Telatar M, Rosenfelt F, Grody W, Territo M, Rosen P.

UCLA Medical School, Division of Hematology/Oncology, Los Angeles, CA 90095, USA. cemmanoednet.ucla.edu

Treatment with the anti-CD20 antibody rituximab prior to stem cell collection may lead to tumor-free stem cell collections in patients with B-cell lymphoma undergoing autologous stem cell transplantation. To test the feasibility of obtaining polymerase chain reaction (PCR)-negative stem cell collection, 30 patients with a variety of B-cell lymphomas were enrolled in a protocol employing a common MINE (mitoxantrone/ifosfamide/etoposide) salvage regimen with rituximab (in vivo purging). Rituximab 400 mg/m2 was administered weekly for 3 weeks on days 1, 6, and 8 in relation to the last MINE cycle, which was followed by growth factor-stimulated peripheral stem cell collection. The median number of CD34(+) cells/kg was 2.5 million cells/kg collected over a median of 5 days. Polymerase chain reaction amplification for the t (14;18) or the heavy-chain gene rearrangement was performed prior to treatment and on the leukapheresis sample. Out of 15 patients who had a positive PCR signal prior to treatment, 10 had PCR-negative stem cell collections, whereas 5 had PCR-positive stem cell collections. After high-dose chemotherapy and stem cell transplant, all patients with a PCR-positive signal pretreatment became PCR negative. We conclude that rituximab may increase the yield of tumor-free stem cells. Higher rates of PCR negativity have been reported when more intense and protracted chemoimmunotherapy regimens have been employed. The magnitude of clinical benefit and the significance of the PCR analysis in stem cells after rituximab requires larger studies.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12435284&dopt=Abstract [PubMed - in process]

J Craniofac Surg. 2002 Mar;13(2):231-9; discussion 240-3.
Tissue-engineered bone using mesenchymal stem cells and a biodegradable scaffold.

Boo JS, Yamada Y, Okazaki Y, Hibino Y, Okada K, Hata K, Yoshikawa T, Sugiura Y, Ueda M.

Department of Oral and Maxillofacial Surgery, Graduate School of Medicine, Nagoya University, 65 Tsuruma-cho, Showa-ku, Nagoya 466-8550, Japan. saiseifed.nagoya-u.ac.jp

Bone marrow has been shown to contain a population of rare cells capable of differentiating to the cells that form various tissues. These cells, referred to as mesenchymal stem cells (MSCs), are capable of forming bone when implanted ectopically in an appropriate scaffold. The aim of this study was to investigate the potential of a new beta-tricalcium phosphate (beta-TCP) as a scaffold and to compare the osteogenic potential between beta-TCP and hydroxyapatite (HA). The beta-TCP and HA loaded with MSCs were implanted in subcutaneous sites and harvested at 1, 2, 4, and 8 weeks after implantation for biochemical and histological analysis. Biochemically, in both beta-TCP and HA composites, the alkaline phosphatase activity in the composites could be detected and was maintained at a high level for 8 weeks. In the histological analysis, active bone formation could be found in both the beta-TCP and HA composites. These findings suggest that beta-TCP could play a role as a scaffold as well as HA. The fabricated synthetic bone using biodegradable beta-TCP as a scaffold in vivo is useful for reconstructing bone, because the scaffold material is absorbed several months after implantation.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12000879&dopt=Abstract

J Biomed Mater Res. 2002 Jul;61(1):75-82.
Chondroinduction of mouse mesenchymal stem cells in three-dimensional highly porous matrix scaffolds.

Aung T, Miyoshi H, Tun T, Ohshima N.

Department of Biomedical Engineering, Institute of Basic Medical Sciences, University of Tsukuba, Ibaraki 305-8575, Japan.

Porous polyvinyl formal (PVF) resin and poly(lactide-caprolactone) [P(LA/CL)] sponges were examined as three-dimensional matrices for chondroinduction of cultured bone marrow mesenchymal stem cells (MSCs). Approximately 5 x 10(5) mouse MSCs were seeded in porous PVF resin or P(LA/CL) sponges and were cultured for up to 1 month in serum-free high-glucose Dulbecco's modified Eagle's medium containing 10 ng/mL transforming growth factor-beta3 and 100 nM dexamethasone for chondroinduction. After the 1-month culture period, the PVF resin and P(LA/CL) sponges contained approximately twice the amount of glycosaminoglycans compared with the control pellet. Safranin-O staining of PVF and P(LA/CL) after 1 month of culture revealed a cartilage-like extracellular matrix containing glycosaminoglycans and collagen. When implanted into nude mice, PVF and P(LA/CL) seeded with MSCs were found to be both biocompatible and chondroinductive. These highly porous scaffolds can maintain a large number of cells in a three-dimensional structure. Both are potentially promising for the chondroinduction of bone marrow MSCs for research and clinical applications. 2002 Wiley Periodicals, Inc.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12001249&dopt=Abstract

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