Neuropeptide W is present in antral G cells of rat, mouse, and human stomach.

Neuropeptide W (NPW) is a 30-amino-acid peptide initially isolated from the porcine hypothalamus as an endogenous ligand for the G protein-coupled receptors GPR7 and GPR8. An intracerebroventricular administration of NPW increased serum prolactin and corticosterone concentrations, decreased dark-phase feeding, raised energy expenditure, and lowered body weight. Peripherally, GPR7 receptors are abundantly expressed throughout the gastrointestinal tract; the presence of NPW in the gastrointestinal endocrine system, however, remains unstudied. Using monoclonal and polyclonal antibodies raised against rat NPW, we studied the localization of NPW in the rat, mouse, and human stomach by light and electron microscopy. NPW-immunoreactive cells were identified within the gastric antral glands in all three species. Double immunohistochemistry and electron-microscopic immunohistochemistry studies in rats demonstrated that NPW is present in antral gastrin (G) cells. NPW immunoreactivity localized to round, intermediate-to-high-density granules in G cells. NPW-immunoreactive cells accounted for 90% chromagranin A- and 85% gastrin-immunoreactive endocrine cells in the rat gastric antral glands. Using reversed-phase HPLC coupled with enzyme immunoassays specific for NPW, we detected NPW30 and its C-terminally truncated form, NPW23, in the gastric mucosa. Plasma NPW concentration of the gastric antrum was significantly higher than that of the systemic vein, suggesting that circulating NPW is derived from the stomach. Plasma NPW concentration of the gastric antrum decreased significantly after 15-h fast and increased after refeeding. This is the first report to clarify the presence of NPW peptide in the stomachs of rats, mice, and humans. In conclusion, NPW is produced in gastric antral G cells; our findings will provide clues to additional mechanisms of the regulation of gastric function by this novel brain/gut peptide.

Extremely high dose neutron dosimetry using CR-39 and atomic force microscopy.

Atomic force microscopy (AFM) has been applied to the analysis of CR-39 nuclear track detectors for high dose neutron dosimetry. As a feasible study to extract the neutron dose, we have employed a (239)Pu-Be neutron source with the traditional track density measurement of recoil proton etch pits from a high density polyethylene (CH(2)) radiator. After very short etching ( approximately 1 microm), etch pit densities were measured as a function of neutron fluence (neutron dose) up to 1.4 x 10(10) cm(-2) (6.6 Sv). Neutron sensitivity was also measured to be 6.6 x 10(-4). Maximum measurable neutron dose was estimated to be approximately 200 Sv by measuring the fraction of the total image area occupied by the etch pits.

Separation of B and T lymphocytes by a hybrid field-flow fractionation/adhesion chromatography technique.

A hybrid of the techniques of cellular adhesion chromatography and field-flow fractionation has been used for the effective separation of rat mesenteric B and T lymphocytes with nearly complete recovery of both cell species. Use of this hybrid technique also allows the relative binding strengths of cells to biomaterials and other surfaces to be rapidly and simply estimated. For the copolymer surface used here, B cells appear to bind with a force five times greater than T cells.

Lethal form of fibuloulnar A/hypoplasia with renal abnormalities.

The present report describes two sibs with lethal acrorenal developmental complex who were products of a nonconsanguinous marriage. Clinical and roentgenologic characteristics did not match any of the known types of fibuloulnar dysostosis. Distinct abnormalities included lethality at birth, facial anomalies, ear anomalies, symmetrical mesomelic shortness of long bones, fibular agenesis, normal vertebrae, oligosyndactyly of phalanges, congenital heart defect, and cystic or hypoplastic kidney. These cases suggest a new lethal form of recessively inherited fibuloulnar dysostosis with renal anomalies.

Separation of B and T lymphocytes by cellular adsorption chromatography with polyamine graft copolymers as column matrices. II. Recovery of adsorbed B cell enriched populations from the column.

Poly(2-hydroxyethyl methacrylate)-graft-polyamine copolymers were developed for column adsorbents for separating lymphocyte subpopulations collected from rat mesenteric lymph nodes. Bead-shaped adsorbents were prepared by coating poly(2-hydroxyethyl methacrylate)-graft-polyamine copolymer on glass beads. Separation features of the column packed with poly(2-hydroxyethyl methacrylate)-graft-polyamine copolymer-coated glass beads were evaluated under various operating conditions. Separation efficacy, AB/AT, was significantly affected by the infusion rate of lymphocyte suspension into the column, increasing with decreasing infusion rate. T cell purity in the column effluent was almost 95% at the infusion rate of 0.1 ml/min. The infusion rate was also found to affect the detachment of adsorbed lymphocytes from the poly(2-hydroxyethyl methacrylate)-graft-polyamine copolymer surface by mechanical pipetting; lymphocytes adsorbed at a low infusion rate were able to be detached quantitatively by suspending the lymphocyte-adsorbed beads with gentle pipetting. The detached lymphocytes thus obtained were confirmed to be enriched in B cells 1.4 times the initial mixture of B and T cells. The addition of albumin in the medium was found to affect the processes of adsorption and recovery of lymphocyte.

Quantitative evaluation of rat lymphocyte adsorption on microdomain structured surfaces of poly(2-hydroxyethyl methacrylate)/polyamine.

The adsorption behaviour of rat lymphocytes on poly(2-hydroxyethyl methacrylate)-graft-polyamine (HA) copolymers was evaluated using a newly developed chromatographic method. The quantity of lymphocyte adsorption can be varied by regulating the polyamine content in the HA copolymer. A remarkable depression in lymphocyte adsorption was observed on the surface of HA copolymer, consisting of 7 wt% of polyamine graft and 93 wt% of poly(2-hydroxyethyl methacrylate) (pHEMA) backbone. Further introduction of a polyamine graft on pHEMA resulted in the increase of lymphocyte retention on the copolymer surfaces. Lymphocytes adsorbed on HA copolymer surface retained their original round shape. Detailed analysis of the chromatogram showed that interaction of lymphocytes with HA copolymer was very much weaker than that with homopolymer of pHEMA or polyamine.

Elimination of cellular active adhesion on microdomain-structured surface of graft-polyamine copolymers.

To investigate the mechanisms involved in lymphocyte adsorption on poly(2-hydroxyethyl methacrylate)-graft-polyamine copolymer, which is utilized as an adsorbent for cell separation, the role of cellular metabolism in lymphocyte adsorption on these copolymers was evaluated. We examined the effect of lowering environmental temperature and of the drug cytochalasin B that inhibits reorganization of microfilaments in cellular cytoskeletons on lymphocyte adsorption. Although the adsorption of lymphocytes on poly(2-hydroxyethyl methacrylate) was considerably reduced in the presence of cytochalasin B or by lowering temperature, no marked influence of these factors was observed for lymphocyte adsorption on poly(2-hydroxyethyl methacrylate)-graft-polyamine copolymers. These results suggest that, in contrast to common plastics surfaces including poly(2-hydroxyethyl methacrylate), the surface of poly(2-hydroxyethyl methacrylate)-graft-polyamine copolymers does not stimulate or activate adsorbed lymphocytes.

Immunoaffinity chromatography of lymphocyte subpopulations using tert-amine derived matrices with adsorbed antibodies.

New polymeric solid-phase matrices for cell affinity chromatography were prepared and their advantageous characteristics compared with conventional matrices were highlighted. These new matrices are derivatives of poly(2-hydroxyethyl methacrylate) (PHEMA) containing a slight quantity of amino compounds as a co-monomer. They were applied to immunoaffinity selection between IgG+ and IgG- lymphocytes of the rat mesenteric lymph node. Simple physical adsorption was sufficient for anti-rat IgG antibodies to be immobilized on these matrices, allowing us to omit the laborious procedure of covalent-linking of antibodies on a matrix. As these matrices themselves showed extremely low non-specific adsorption of lymphocytes, a very dilute solution of antibody (0.02-0.08 mg/ml) was enough for column conditioning. This separation method gave IgG- lymphocytes of more than 90% purity and almost 95% yield within as short a time as 7 min. Further, IgG+ lymphocytes were obtained in good yield (80-90% of loaded number) by recovering the adsorbing cell fraction from the column by gentle pipetting of the matrix.

Effect of microphase separated structure of polystyrene/polyamine graft copolymer on adhering rat platelets in vitro.

Rat platelet adhesion on microphase separated surface of polystyrene/polyamine graft copolymers (SA copolymers) was investigated. Shapes of adhering platelet were very much changeable in response to the mode of microphase separation of SA copolymer surfaces. We assume that the microphase separated structure of SA copolyer may regulate the shape change of platelet through its effect on a redistribution of proteins and/or lipids present at the plasma membrane of platelets.

Recombinant human glycosylated granulocyte colony-stimulating factor (rhG-CSF)-combined regimen for allogeneic bone marrow transplantation in refractory acute myeloid leukemia.

Recombinant human glycosylated G-CSF (rhG-CSF) may stimulate proliferation of myeloid leukemia cells and thereby increase their susceptibility to anti-cancer agents. By in vitro colony assay, the rhG-CSF-responsive NFS-60 leukemic cell clones are more effectively killed by Ara C in the presence of rhG-CSF than in the absence of rhG-CSF, while the killing of the rhG-CSF-unresponsive HL-60 cell clones is unaffected by rhG-CSF. Leukemia cell colony forming units (L-CFU) derived from most AML patients demonstrate similar results to those of the NFS-60 cell clone when treated in vitro. Encouraged by these in vitro results, we used rhG-CSF as a component of a conditioning regimen for 15 relapsed AML patients who were receiving allogeneic BMT. The patients were conditioned with total body irradiation (TBI) and high-dose Ara C. rhG-CSF was infused continuously at a dose of 5 micrograms/kg/day from 24 h before the beginning of TBI to the end of Ara C therapy. Proliferation of the leukemia cells in vivo in response to rhG-CSF was confirmed in 7 of 14 patients tested and the combined use of rhG-CSF had no additional adverse effects. After BMT, four patients died of non-leukemic causes and three patients had leukemic relapse: the other eight patients have remained disease-free for 200-1600 (median 417) days. The actuarial probabilities of relapse and disease-free survival (DFS) at 4.4 years after BMT were 43.2% and 41.7%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Successful treatment of relapsed blastic natural killer cell lymphoma with unrelated cord blood transplantation.

The prognosis for blastic natural killer (NK) cell lymphoma is generally dismal. We report a patient who was successfully treated with unrelated cord blood transplantation (UCBT). A 15-year-old boy was diagnosed as having blastic NK cell lymphoma in the cervical lymph nodes. Autologous peripheral blood stem cell transplantation was performed on achieving a complete remission. However, the disease recurred in the bone marrow 6 months later. Chemotherapy induced a second remission and the patient received UCBT with a conditioning regimen consisting of total body irradiation, thiotepa and cyclophosphamide. Chronic GVHD of the lung occurred, but it was well controlled with steroids. At the time of writing, he remains in remission 18 months after UCBT with an excellent performance status. UCBT may be an option for patients with blastic NK cell lymphoma.

Human cytomegalovirus infection in foci of Langerhans cell histiocytosis.

Langerhans cell histiocytosis (LCH) has been thought to be a disorder of immune regulation, and increasingly, evidence showing that the tissue damage in LCH involves lymphokines and pro-inflammatory cytokines is reported. We detected human cytomegalovirus (HCMV)-DNA in LCH cells in the foci of LCH lesions by immunohistochemistry, in situ hybridization and PCR. HCMV was detected in the nuclei and/or cytoplasm of LCH cells in 9 of 27 LCH cases by immunostaining. HCMV was probably an early antigen. In situ hybridization revealed signals for HCMV-DNA only in the nuclei of LCH cells in 10 of the 27 LCH cases. PCR analysis was performed in 20 of the LCH cases, and HCMV-DNA was detected in 7 of these. All 7 positive cases were also positive for HCMV by ISH and IHC. These findings suggested that early phase infection or reactivation of HCMV occurred in the LCH lesions. HCMV infection may be accompanied by impaired cytokine production. Our study also suggested a relationship between HCMV infection and expression of TNFalpha. In tissues affected by LCH, dermatopathic lymphadenopathy or malignant fibrous histiocytoma and in normal tissues no signals for Epstein-Barr virus-RNA were detected. These findings suggest that in some cases LCH is associated with HCMV infection.

Effects of glycerol on intracellular ice formation and dehydration of onion epidermis.

Microscopic observations of freezing processes of onion epidermis have been carried out using a directional solidification stage. Effects of cryoprotectant on the intracellular ice formation and the dehydration were examined for several kinds of concentrations of glycerol in a wide range of cooling rate. With an increase of the glycerol concentration the ice formation due to the surface-catalyzed nucleation (SCN) at the plasma membrane was suppressed and the freezing temperature decreased. Especially, under a rapid cooling condition, a large supercooling should be achieved and we observed a different type of freezing in the lower temperature conditions. At the slower cooling rate, on the other hand, the use of glycerol can not prevent the cell shrinkage caused by the dehydration. It is also found that the intracellular ice formation and the dehydration under the ordinary cooling condition can be estimated based on the SCN theory by Toner et al. and Mazuar's dehydration model.

Alkaline phosphatase, defensin gene expression and effect of myeloid cell growth factors in normal and leukemic cells.

In neutrophils, four different granules are defined, i.e. azurophil, specific, gelatinase and secretary vesicles. In these granules many neutrophil-specific constituents are identified. Some of these constituents have already been cloned and their gene expressions studied. In such constituents, alkaline phosphatase (ALP) and defensin are well known, although their functions are not yet fully clarified. ALP is present in secretary vesicles and has important roles in the diagnosis of some myeloid disorders. On the other hand, defensin is the most abundant functional peptide of neutrophils and is present in azurophillic granules. Which are subdivided into defensin-rich and defensin-poor granules. This review describes the expression of ALP and the defensin gene in normal and leukemic cells and the effect on these genes of myeloid growth factors, such as granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor and interleukin 3.

Myeloperoxidase gene expression and regulation by myeloid cell growth factors in normal and leukemic cells.

Myeloperoxidase (MPO) is present in azurophilic granules which appear in the promyelocyte stage of differentiation, and is the most common functional protein of myeloid cells. With progress in molecular biology, the expression and regulation of MPO have been clarified in normal myeloid and leukemic cells, not only by enzymatical activity but at the gene level MPO expression is affected by the differentiation of myeloid cells, and has been suggested to be regulated by myeloid cell growth factors, such as granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor and interleukin-3. In the past decade the signal transduction from their receptors has been clarified. This review describes the expression and regulation of the MPO gene in myeloid cells including myeloid disorders, such as myeloid leukemia or myelodysplastic syndromes, The effects on MPO by myeloid growth factors and signal transduction from their receptors are also presented.

In vivo release of cholecystokinin-like immunoreactivity in rat frontal cortex under freely moving conditions.

Release of cholecystokinin-like immunoreactivity (CCK-LI) was measured in rat medial prefrontal cortex in vivo by brain dialysis and enzyme immunoassay under freely moving condition. Perfusion of 50 mM K+ resulted in the increase of CCK-LI in the dialysate. The data on high-performance liquid chromatography (HPLC) of the dialysate showed that the increase of the CCK-LI was mainly due to the increase of CCK octapeptide sulfate itself. In the preliminary experiments, we applied this brain dialysis method for determination of CCK-LI release with drug treatment. After treatment with sulpiride (i.p.), a D2 dopamine receptor antagonist, a significant increase of CCK-LI was observed, indicating that this brain dialysis technique is applicable to detect change in the level of CCK-LI release after a certain drug treatment.

Mycobacterium kansasii arthritis of the foot in a patient with systemic lupus erythematosus.

Mycobacterium kansasii, an atypical Mycobacterium, may cause serious illness in humans. We describe a M. kansasii infection of the foot joint, which was diagnosed in a 46-year-old woman with systemic lupus erythematosus. The diagnosis was based on a positive culture from degenerative tissue and histological diagnosis of a synovium. We reviewed the literature regarding M. kansasii infection of the joint, bone, and periarticular structures focusing on the complication of rheumatic diseases.

Functional outcome of anterior transposition of the vascularized ulnar nerve for cubital tunnel syndrome.

Anterior transposition of the ulnar nerve is a widely used treatment for cubital tunnel syndrome, but neurolysis performed at the time of surgery may impair the blood supply to the ulnar nerve. This study compared the results of intramuscular anterior transposition of the ulnar nerve with or without preserving the extrinsic vessels of the ulnar nerve in 35 patients. The postoperative nerve conduction velocity and the clinical results were better in the group in which the extrinsic vessels were presented.