Overexpression of C-terminal Src kinase homologous kinase suppresses activation of Lyn tyrosine kinase required for VLA5-mediated Dami cell spreading.

The Csk homologous kinase (Chk), which is co-expressed with C-terminal Src kinase (Csk) in hematopoietic cells, negatively regulates Src family kinases in vitro with selectivity toward Lyn but not c-Src in platelets. To explore the role of Src family kinases in hematopoietic cell adhesion, we overexpressed Chk in the megakaryocytic cell line Dami and established clones exhibiting a 10-fold increase in the amount of Chk. Overexpression of Chk was found to suppress VLA5 integrin-mediated cell spreading, but not cell attachment, throughout fibronectin (FN) stimulation.

Translocation of the Csk homologous kinase (Chk/Hyl) controls activity of CD36-anchored Lyn tyrosine kinase in thrombin-stimulated platelets.

Chk/Hyl is a recently isolated non-receptor tyrosine kinase with greatest homology to a ubiquitous negative regulator of Src family kinases, Csk. To understand the significance of co-expression of Chk and Csk in platelets, we examined the subcellular localization of each protein. Chk, but not Csk, was completely translocated from the Triton X-100-soluble to the Triton X-100-insoluble cytoskeletal fraction within 10 s of thrombin stimulation.

Analysis of CSK homologous kinase (CHK/HYL) in hematopoiesis by utilizing gene knockout mice.

CHK/HYL is a non-receptor tyrosine kinase that belongs to CSK (C-terminal Src kinase) family. Northern blotting and RT-PCR analyses showed that CHK/HYL was expressed in large spectrum of hematopoietic cells except for erythroid cells and brain. To explore the function of CHK/HYL in hematopoietic cells, we generated CHK/HYL deficient mice.

The role of olfaction in oil preference in the chicken.

The role of olfaction on the preference of diets containing 20% medium-chain (MCT) or long-chain triacylglycerol (LCT) was investigated in the chicken. Olfactory bulbectomized, sham-operated (Sham) or intact (Intact) birds were offered a choice between LCT or MCT diet and food intake was measured over a short time period. Intact and Sham groups showed a significant preference for LCT over MCT diet, but olfactory-bulbectomized chickens lost the preference for LCT over MCT.

Polysynaptic pathways from high threshold muscle afferents innervating hindlimb muscles to tail motoneurons in the spinalized cat.

The postsynaptic potentials (PSPs) after stimulating hindlimb flexor (PBSt) and extensor muscle nerves (LGS) were recorded from 64 tail motoneurons innervating m. extensor caudae lateralis and m. flexor caudae longus in the spinalized cats.

Diagnosis of premature rupture of membranes with an improved alpha-fetoprotein monoclonal antibody kit.

We developed a new kit for detecting alpha-fetoprotein (AFP) in leaked amniotic fluid (Eur J Obstet Gynecol Reprod Biol 1995;58:67-72). Later, we developed an improved AFP kit utilizing the same AFP monoclonal antibody. We compared this improved AFP test with the nitrazine test for 137 patients.

Quantitative and qualitative comparative analysis of gradient-separated hematopoietic cells from cord blood and chemotherapy-mobilized peripheral blood.

Some of the uncertainty regarding the use of cord blood (CB) in transplant settings includes the suspected relative rarity of hematopoietic stem cells (SC) in CB and the feasibility of incorporating a cell separation protocol to remove red blood cells, which may result in an unacceptable loss of SC. To address this uncertainty, we isolated a SC fraction by Percoll or Ficoll gradients from CB and peripheral blood (PB), which had been mobilized by chemotherapy. The frequencies of mononuclear cells (MNC), CD34+ cells, colony-forming units granulocyte-macrophage (CFU-GM), and the output of colony-forming cells (CFC) after five weeks in long-term culture (LTC) assay were then evaluated.

Polysynaptic pathways from hindlimb cutaneous afferent nerves to tail muscle motoneurons in unanesthetized and spinalized cats.

Postsynaptic potentials (PSPs) were recorded in 57 extensor caudae lateralis (ECL) motoneurons and 82 flexor caudae longus (FCL) motoneurons of 36 decerebrated and spinalized (L1-2) adult cats, upon electrical stimulation of hindlimb cutaneous nerves, such as the sural nerve, superior peroneus nerve, and plantar section of the tibial nerve. With low threshold stimulation (1.2-5.

Effects of hindlimb cutaneous afferent inputs on spinal reflex recording from tail muscle motoneurons in the spinalized cat.

In 4 spinalized cats, the effects of afferent inputs from hindlimb cutaneous nerves (sural cutaneous nerve: Sur) on mono-and poly-synaptic reflex recorded from tail muscle motoneurons were studied before and after spinal lesioning at S2-3 level. Monosynaptic reflex was enhanced by ipsilateral Sur stimulation at short conditioning-test stimulus interval and this effect was not observed after spinal lesion of ipsilateral side of spinal cord. Polysynaptic reflex was inhibited by stimulation of Sur in both sides and this inhibitory effect was depressed by contralateral spinal lesioning.

Effects of monocyte-macrophage colony-stimulating factor (M-CSF) on in vitro erythropoiesis of marrow progenitor cells from patients with renal anemia.

We examined the influence of monocyte-macrophage colony-stimulating factor (M-CSF) on erythropoiesis both in vitro and in vivo in 98 patients with chronic renal failure who were undergoing hemodialysis. Serum levels of M-CSF and the clinical response to therapy with human recombinant erythropoietin (Epo) were analyzed. The following results were obtained: 1) The serum level of M-CSF was 6.

Optimal growth of human blood hematopoietic progenitor cells collected by apheresis for autografts.

For safe autografts with peripheral blood hematopoietic cells (PBSCT), better methods for determining the kinetics of stem cell populations and predicting engraftment speed after PBSCT need to be established. Current methods include culture in semi-solid medium and measurement of CD34 cell surface antigen. In this study with only partially purified blood cells obtained from children with cancer in remission, we compared the effects of phytohemagglutinin-stimulated lymphocyte-conditioned medium (PHA-LCM) and recombinant human cytokines on the growth of progenitor cells in a methylcellulose culture system.

Synergism of interleukin 12, interleukin 3 and serum factor on primitive human hematopoietic progenitor cells.

Interleukin 12 (IL-12), or natural killer cell stimulatory factor (NKSF), has multiple biologic effects on T lymphocytes and natural killer cells. In this study, we evaluated the effect of IL-12 on human hematopoiesis by analyzing the growth of CD34+33- cells in methylcellulose culture with or without serum. Blood cells were collected by apheresis from an expanded stem cell pool during the rebound phase of hematopoiesis after chemotherapy and purified by combined purification procedures to yield target CD34+33- cells.

Synergy among erythropoietin, interleukin 3, stem cell factor (c-kit ligand) and interferon-gamma on early human hematopoiesis.

The stimulatory effect of interferon-gamma (IFN-gamma) on human early hematopoiesis was investigated using CD34+ cells that were purified from peripheral blood as a target. In the presence of human interleukin 3 (IL-3) and stem cell factor (SCF), the growth of colony forming units for granulocyte/macrophage (CFU-GM) in serum-free methylcellulose culture was enhanced up to 5.5-fold over baseline colony formation (n = 10, mean +/- SE, 3.

Regeneration of immunity and varicella-zoster virus infection after high-dose chemotherapy and peripheral blood stem cell autografts in children.

We assessed recovery of the immune system in 41 children who underwent high-dose chemotherapy (without total body irradiation) and autologous peripheral blood stem cell transplantation (PBSCT) for acute leukemias or non-Hodgkin's lymphoma. The analysis was in two parts. Firstly, we performed serial monitoring of regenerating subsets and blastogenesis of lymphocytes.

Engraftment potential of peripheral and cord blood stem cells evaluated by a long-term culture system.

The experiment was performed in an attempt to seek a suitable indicator of allogeneic transplantation using cord blood cells. The number of colony-forming cells (CFC) recovered after 5 weeks of culture represents that of primitive progenitors present in the initiating cell population. In this study of nine children who underwent autologous peripheral blood stem cell transplantation (PBSCT), we measured the amount of colony-forming units-granulocyte/macrophage (CFU-GM) and total CFC output after 5 weeks in long-term culture (LTC-CFC) contained in the infused grafts and compared them to the speed of hematopoietic recovery after marrow ablation.

Comparative analysis of engraftment after cryopreservation of peripheral blood stem cell autografts by controlled- versus uncontrolled-rate methods.

Peripheral blood stem/progenitor cells (PBSC) were cryopreserved by different methods and their clonogenic viabilities were compared. The traditional cryopreservation method involves controlled-rate freezing with 10% dimethyl sulfoxide (DMSO) and a programmed freezer (PF). The alternative method incorporates 6% hydroxyethyl starch and 5% DMSO without PF.

Toxicities associated with cryopreserved and thawed peripheral blood stem cell autografts in children with active cancer.

To evaluate the safety of cryopreserved and thawed peripheral blood stem cell (PBSC) autografts in children with active cancer, a toxicity assessment was made of 54 PBSC transfusions to 52 children (aged 1-16 years; median, 9 years). Patients were conditioned with high-dose chemotherapy without total body irradiation. The volume of PBSCs transfused varied from 46 to 500 mL (219.

Effects of immunosuppressants, FK506, deoxyspergualin, and cyclosporine A on immature human hematopoiesis.

Effects of the immunosuppressants, FK506, deoxyspergualin (DSG), and cyclosporine A (CsA) on the growth of human hematopoietic progenitor cells were tested in the presence of interleukin-3 (IL-3) with purified bone marrow and blood cells as targets in methylcellulose culture. FK506 had a significant stimulatory effect on the growth of colony-forming units/granulocyte-macrophage (CFU-GM) and burst-forming units/erythroid (BFU-E) from peripheral blood and cord blood cells but not from bone marrow cells. Neither DSG nor CsA had an effect on any type of target cell.

Measurement of serum levels of macrophage colony-stimulating factor (M-CSF) in patients with uremia.

Serum levels of monokines, including macrophage colony-stimulating factor (M-CSF), tumor necrosis factor-alpha (TNF-alpha), interleukin-1 alpha (IL-1 alpha) and IL-1 beta (IL-1 beta), were measured in patients with chronic renal failure in an attempt to clarify the kinetics of these cytokines in the course of renal anemia. M-CSF was the only monokine detectable in the serum from all patients as well as healthy donors, making this cytokine feasible and reliable for serial evaluations. On all occasions, the level of M-CSF in uremic patients was significantly higher than that in healthy donors (29.

Granulocyte colony-stimulating factor (CSF), macrophage-CSF, granulocyte-macrophage CSF, interleukin-3, and interleukin-6 levels in sera from children undergoing blood stem cell autografts.

Endogenous production of granulocyte colony-stimulating factor (G-CSF), macrophage CSF (M-CSF), granulocyte-macrophage CSF (GM-CSF), interleukin-3 (IL-3), and interleukin-6 (IL-6) was investigated in 10 children who underwent a total of 12 courses of autologous peripheral blood stem cell transplant (PBSCT) by measuring their serum levels using immunoassay kits. The serum G-CSF level increased immediately following infusion of PBSC graft, peaked between days 3 and 7 posttransplant and then declined by the time the granulocyte count rose. No definitive association was found between the continuous high levels of G-CSF and infective episodes, the number of infused nucleated cells, monocytes, CFU-GM, or the number of days required to achieve greater than 0.

[Evaluation of the benefits of sodium 2-mercaptoethane sulfonate (MESNA) therapy for children undergoing high-dose chemotherapy].

Preventive effects of hemorrhagic cystitis by the use of sodium 2-mercaptoethane sulfonate (MESNA) was evaluated in 40 children undergoing peripheral blood stem cell autografts (PBSCT) after marrow-ablative chemotherapy which included high-dose cyclophosphamide (CY, 50mg/kg x 2). Fifteen patients received MESNA (group A) and 25 did not (group B), and all received concomitant hyperhydration (3,000ml/m2/day). No renal dysfunction or toxicity attributed to the use of MESNA was observed in either group of patients.