Effects of cytokines on hematopoietic progenitor cells in cord blood,in bone marrow,and in peripheral blood mobilized by chemotherapy and G-CSF

We compared the effects of various combinations of cytokines (stem cell factor [SCF], interleukin [IL] ?3, IL-6, granulocyte-colony stimulating factor [G-CSF], erythropoietin [EPO]) among the growth of human hematopoietic progenitor cells from cord blood (CB), bone marrow (BM), and peripheral blood mononuclear cells (MNC) mobilized by chemotherapy and G-CSF (PB) in a semi-solid medium. Macroscopic colonies, that were visible to the naked eye, were formed from PB-MNC within 1 week even without cytokines. They consisted of blasts containing macrophage-like cells with immature nuclei on Wright stain, and were strongly accelerated by IL-3. Macroscopic colonies were also formed from CB-MNC. However, they appeared after 1–3 weeks and synergistic effects of SCF with other cytokines, especially EPO, were prominent. Macroscopic colonies were not formed from BM-MNC. Granulocyte-colony stimulating factor was effective in increasing colony forming units of granulocyte macrophage from BM-MNC and they appeared between 1 and 2 weeks. These results suggested that the quality of hematopoietic progenitor cells was different among blood sources. This might lead to different bone marrow recovery patterns after transplantation of each blood source. The appropriate cytokines should be added to evaluate their exact potential.     

Ex vivo expansion of umbilical cord blood hematopoietic progenitor cells by combinations of cytokines

Ex vivo expansion of hematopoietic progenitor cells in the umbilical cord blood mononuclear cells (CB-MNC) was investigated in liquid culture system with various combinations of cytokines (stem cell factor [SCF], interleukin [IL]-3, IL-6, granulocyte-colony stimulating factor [G-CSF], erythropoietin [EPO], and interferon [INF]-γ). Non-lineage-committed hematopoietic progenitor cells and lineage committed hematopoietic progenitor cells were represented as CD34⁺CD38^? and CD34⁺CD38⁺ subpopulations, respectively. Although absolute CD34⁺CD38^? cell numbers decreased even in the presence of multicytokines, the combinations of SCF plus IL-6 and SCF plus IL-3 plus IL-6 plus INF-γ were significantly effective in maintaining CD34⁺CD38^? cells than the other combinations (P < 0.05). After 4 weeks of culture, CD34⁺CD38^? cells disappeared in all combinations of cytokines. Absolute CD34⁺CD38⁺ cell numbers increased in the presence of cytokines. Maximal expansion of CD34⁺CD38⁺ cells were observed in the combinations of SCF plus IL-3 plus IL-6 plus EPO (19.8 ± 3.3 -fold) and SCF plus IL-3 plus IL-6 plus G-CSF (18.3 ± 2.6). The combination of SCF plus IL-3 plus IL-6 was also effective to expand CD34⁺CD38⁺ cells (15.8 ± 3.9). However, the expansion was transient and they decreased to zero within 3 weeks. In the combinations of SCF plus IL-6 and SCF plus IL-3 plus IL-6 plus INF-γ, maximal expansion was inferior to the others but CD34⁺CD38⁺ cells were maintained more than 4 weeks. These results suggested that the indication of CBT can be expanded into older children by ex vivo augmentation of CB hematopoietic progenitor cells using multi-cytokines.     

Prediction of engraftment after peripheral blood stem cell transplantation by CD34-positive cells in grafts

total of 91 peripheral blood stem cell collections were performed in 26 children with various malignant tumors and peripheral blood stem cell transplantations (PBSCT) were performed in 15 of the children. There was a positive correlation between logarithm of total CD34⁺ cells/kg and logarithm of colony-forming unit-granulocyte macrophage (CFU-GM)/kg (r = 0.86). The time elapsed until the white blood cells (WBC) exceeded 1000/μL was related to both CFU-GM (r = 0.67) and CD34⁺ cell count (r = 0.60). The number of days elapsed until platelet count exceeded 5 times 10⁴/μL was not related to the logarithm of CFU-GM count/10⁵ per kg transfused (r = 0.47), but was related to the logarithm of CD34⁺ cell counts/10⁶ per kg transfused (r = 0.73). The number of days elapsed until the reticulocytes exceeded 5 times 10⁴/μL was not related to the logarithm of CFU-GM count/10⁵ per kg (r = 0.52), but was related to the logarithm of CD34⁺ cell counts/10⁶ per kg transfused (r = 0.91). Although CD34⁺ cell counts correlated with the number of CFU-GM, bone marrow regeneration rates in three lineages were predicted more accurately by the number of CD34⁺ cells transfused than by the number of CFU-GM. These results suggest that measurement of the CD34⁺ cell count may be useful in predicting bone marrow regeneration rate after PBSCT.     

Associations of workplace bullying and harassment with stress reactions: a two-year follow-up study

The purpose of this prospective study was to investigate the effect of the patterning of workplace bullying and harassment over two time points (chronic, remission, onset, and never) on psychological and physical stress reactions. The subjects were 543 workers at welfare facilities for the elderly in Japan who completed a self-administered questionnaire at Time 1 (from August to September, 2009) and at Time 2 (from September to October, 2011). Workplace bullying and harassment were assessed using the Negative Acts Questionnaire (NAQ). Stress reactions were assessed using the Brief Job Stress Questionnaire. In the multiple logistic regression analyses, onset of person-related bullying was significantly (p<0.05) positively associated with both psychological and physical stress reactions at Time 2. Chronic form of person-related bullying was significantly (p<0.05) positively associated with psychological stress reaction at Time 2. Onset of sexual harassment was significantly (p<0.05) positively, and remission of sexual harassment was significantly (p<0.05) negatively associated with physical stress reaction at Time 2. Onset and chronic form of person-related bullying and onset of sexual harassment can cause stress reactions. Remission of sexual harassment can terminate physical stress reaction.     

Physical and socioeconomic predictors of birthweight in Japan

BACKGROUND: The purpose of this study was to characterize physical and socioeconomic factors associated with birthweight in Tokyo, Japan. METHODS: In a cohort of obstetric outpatients from the Tokyo Metropolitan Maternity and Child Health Institute, Tokyo, and The Jikei University School of Medicine, Chiba, Japan, 553 eligible women in their third trimester of pregnancy were given a survey investigating physical and socioeconomic backgrounds. RESULTS: Infants tended to be small for gestational age when maternal pre-pregnancy weight was lower, and low income and female infant gender were significantly associated with a low birthweight infant. Further, birthweight compared with predicted birthweight was lower in the case of a female infant gender, low income, low maternal pre-pregnancy weight and low parity. CONCLUSION: Income is a novel parameter that is associated with birthweight in Japan.     

Nephrotoxicity of a New Cephalosporin, DQ-2556, in Rats

A single intravenous administration of a new cephalosporin,DQ-2556, at 1200 mg/kg to Sprague-Dawley rats induced proximaltubular necrosis. The histological lesions were not closelycorrelated with renal cortical concentrations of DQ-2556. Developmentof renal injuries was examined histologically. The arcuate andcortical radial arteries of the kidneys were constricted immediatelyalter the injection, but then became dilated and showed histologicalchanges: penetration by erythrocytes, edematous thickening,and necrosis in the media. In addition to congestion in theouter medulla, the proximal convoluted and straight tubulesexhibited the earliest changes 30 mm after the injection, namely,enlargement and rounding of the mitochondria and swelling andirregular arrangement of the microvilli in the epithelial cells.Small necrotic foci of epithelium were observed from 4 hr. Theywere mainly distributed in the outer cortex and the outer stripeof the outer medulla 24 hr later. In the examination of hemodynamics,DQ-2556 significantly decreased the blood flow with increasedvascular resistance in the renal artery during and after a singleinjection. These changes had disappeared wholly or partially60 mm after dosing commenced. Fur thermore, Ca²⁺ channel blockersmarkedly inhibited increases in the serum urea nitrogen andcreatinine concentrations and development of the tubular necrosisand the lesions of the arterial walls, which were induced byDQ-2556. These results suggest a possible contribution of theconstriction of renal artery to the tubular necrosis.     

Mucin-Producing Carcinoma of the Thyroid Gland--A Case Report and Review of the Literature

Mucin-producing carcinoma of the thyroid is rare and its histogenesishas been debated. The first case was reported by Diaz-Perezet al. (1976). We herein report the sixth case of mucin-producingcarcinoma of the thyroid which was discovered in a 72-year-oldJapanese woman. This patient was admitted with a 10-year historyof a lump in the right anterior neck, which started to growsuddenly two months before diagnosis. The pathological diagnosisat biopsy was anaplastic carcinoma. A course of radiation therapyfollowed by chemotherapy resulted in no response. She died oftwo months after admission. At autopsy, the main tumor was foundin the right lobe of the thyroid with metastasis to the lungsand liver. Histologically, the tumor was composed of mucin-secretingglandular cells and nests of epidermoid cells with keratinizationand infrequent formation of intercellular bridges. Mucin wasdemonstrated both intracellulary and extracellulary.     

Spermatogenic Disturbance Induced in Mice by Combined Local Injection of Monoclonal Antibodies to Sertoli Cell and to Basal Lamina of Seminiferous Tubule

ABSTRACT: Two kinds of hybridoma clones, one producing monoclonal antibodies against Sertoli cell (TM-1) and the other the basal lamina of the seminiferous tubule (TM-2), were raised by fusion between P3X63Ag8-653 mouse myeloma cells and spleen cells of BALB/c mice (H-2^d) immunized with testicular homogenate of the same inbred mice. Immunohistochemically, TM-1 reacted specifically with cytoplasmic component of Sertoli cell and TM-2 with basal lamina of the seminiferous tubule. Using these monoclonal antibodies, spermatogenic disturbance was induced experimentally in BDF₁(H-2^b/d) by intratesticular injection of a set of these two antibodies. Single injection of either TM-1 or TM-2 failed to induce the lesion. This fact indicated that TM-1 antibody could reach the Sertoli cell to impair its function, which was otherwise inaccessible without coincidental action of TM-2 antibody. TM-2 antibody appeared to alter the permeability of the basal lamina of the tubule and lower its barrier effect.     

Antipsoriatic and antimetabolic agents as stimulators of the beta-adrenergic adenylate cyclase response of epidermis

Using pig skin organ culture in vitro, we compared the effects of antipsoriatic and anti-metabolic agents on the beta-adrenergic response of epidermis. After a 24-h incubation with these chemicals, the beta-adrenergic response of epidermis was shown to be increased to various degrees. Among the chemicals, colchicine, actinomycin D, and puromycin had the most marked effect, and were followed by Ro 10–1670 and hydrocortisone, and then by anthralin, cycloheximide and N-5’(N-(3,4-dimethyloxycinnamoyl) anthranilic acid). Methotrexate and hydroxyurea apparently had no (or rather decreasing) effects on the beta-adrenergic response of epidermis. After the same treatment in vitro, thymidine incorporation of epidermal keratinocytes was compared. In the latter system, the inhibitory effects of puromycin, cycloheximide and actinomycin D were most marked, and were followed by those of colchicine and Ro ro-1670, and then hydrocortisone and anthralin. n-s' had no effect and hydroxyurea increased thymidine incorporation of keratinocytes. Our data indicate that most (but not all) anti-psoriatic agents do augment the beta-adrenergic response of epidermis, which might be associated with the clinical efficacy of these agents in psoriasis where a defective beta-adrenergic response is well documented. The augmentation effects of these chemicals, however, did not correlate with their inhibitory effects on keratinocyte proliferation, at least in the thymidine incorporation system in epidermal keratinocytes in vitro.     

Dilated cardiomyopathy in a patient receiving continuous ambulatory peritoneal dialysis

We describe a unique case of a 10 year old girl with dilated cardiomyopathy who has been receiving continuous ambulatory peritoneal dialysis. She responded to treatment with a β₁-agonist. We note that electrocardiography and echocardiography are effective for early recognition of the onset of dilated cardiomyopathy and that a thallium scan should be performed to assess the severity of dilated cardiomyopathy in patients with chronic renal insufficiency.