Measurement of secondary colony formation after 5 weeks in long-term cultures in patients with myelodysplastic syndrome

Pancytopenia is a frequent manifestation of myelodysplastic syndromes (MDS). In the presence of an empty bone marrow, clinical distinction from aplastic anemia may be difficult. The hypoplastic marrow morphology seen in some cases of MDS raises questions about etiologic and pathophysiologic relationships between aplastic anemia and MDS. The goal of our study was to compare the degree of the hematopoietic failure in these diseases at the level of the most immature progenitor and stem cells that can be measured in vitro. In a systemic, prospective fashion, we have studied bone marrow (n = 45) and peripheral blood (n = 33) of patients with MDS for the number of long-term culture initiating cells (LTC-IC) in comparison to 17 normal controls and patients with new, untreated aplastic anemia (46 marrow; 62 blood samples). Due to the low numbers of cells available for the analysis, formal limiting dilution analysis could not be performed, instead secondary colony-forming cells (CFC) after 5 weeks of LTBMC were measured. As the number of these cells is proportional to the input number of LTC-IC, the number of secondary CFC per 10(6) mononuclear cells (MNC) initiating the LTBMC can be used as a measure of the content of immature stem cells in bone marrow and peripheral blood. The MDS group consisted of 34 RA, three RARS, eight RAEB and two RAEB-T patients with mean absolute neutrophil values of 1992, 1413, 1441, and 380 per mm3, respectively. The diagnosis was established based on bone marrow morphology and results of cytogenetic studies. In comparison to controls (147 +/- 38/10(6) MNC), significantly decreased numbers of bone marrow secondary CFC were found in MDS: in patients with RA and RARS, 21 +/- 7 secondary CFC per 10(6) bone marrow MNC (P < 0.001); patients with RAEB and RAEB-T: 39 +/- 12 CFC per 10(6) marrow MNC (P < 0.001). In all groups tested, the decrease in peripheral blood secondary CFC numbers was consistently less pronounced. In MDS patients with hypocellular bone marrow, secondary CFC were lower but not significantly different in comparison to MDS with hypercellular marrow (18 +/- 6 vs 35 +/- 11; NS; hypoplastic bone marrow also was not associated with significantly lower neutrophil counts). However, in 24% of patients with MDS, bone marrow secondary CFC were within the normal range, while in the aplastic anemia group only one of the patients showed secondary CFC number within normal range. Bone marrow and blood secondary CFC numbers in hypoplastic RA were significantly higher than those in severe aplastic anemia 919 +/- 5 in bone marrow, P < 0.01; 7 +/- 2 in blood, P < 0.05). This trend was even more pronounced in hypoplastic RA with chromosomal abnormalities. However, no significant differences were found between the secondary CFC numbers in hypoplastic RA and moderate aplastic anemia. We concluded that, although the deficiency in the stem cell compartment is less severe in MDS than in aplastic anemia, depletion of early hematopoietic cells is an essential part of the pathophysiology in both diseases.     

Transcriptional analysis of Marek''s disease virus (MDV) genes in MDV-transformed lymphoblastoid cell lines without MDV-activated cells

Prevention of the occurrence of diabetes-specific vascular complications is the final aim of clinical islet transplantation. Pancreatic islets isolated from adult pigs may be a suitable tissue source to transplant a large number of type 1 diabetic patients. Acute cellular rejection may be finally overcome by clinically applicable protocols for tolerance induction. However, primary nonfunction of the graft, as regularly observed in the porcine islet-to-rat xenotransplantation model, may be an additional problem. In this paper, species-specific inflammatory and immunological mechanisms are discussed which prevent early porcine islet graft function in rats but not in mice.     

Direct fractionation of proteins in particle-containing feedstocks by a filter paper pieces-based DEAE-cellulose column chromatography. Rapid, robust and low-cost capturing procedure for protein

Goldfish were classically conditioned to a mixture of two pulse trains differing in both repetition rate and the spectral profile of the pulses. Animals were then tested for generalization to single pulse trains having one or the other spectral profile presented at a variety of repetition rates. Generalization functions of repetition rate were qualitatively similar to those obtained following conditioning to either of the pulse trains alone. Thus, the spectral profile of each pulse type was appropriately associated with the repetition rate at which that pulse type was presented during conditioning. These results indicate that the two concurrent pulse trains making up the conditioning stimuli were analyzed independently, forming two auditory streams. When either of the two pulse trains were presented with a 500 ms onset asynchrony, stream segregation was enhanced. These and other results suggest that many fundamental features of the human sense of hearing are widely shared among vertebrate animals, and may have developed first among fishes.     

Adaptive functioning following traumatic brain injury and orthopedic injury: a controlled study

BACKGROUND: In an effort to intensify osteosarcoma therapy, systemic ifosfamide was added pre- and postoperatively to an already aggressive three-drug regimen. In a subgroup of patients, loco-regional treatment intensification was attempted by using the intraarterial route to give cisplatin. PATIENTS AND METHODS: Patients < or = 40 years at diagnosis of a localised, de novo high-grade central extremity osteosarcoma were eligible for inclusion into study COSS-86 if registered within three weeks from biopsy. Doxorubicin, high-dose methotrexate, and cisplatin were given to all patients. Patients who fulfilled one or more of three defined high-risk criteria received early systemic treatment intensification by adding ifosfamide as the fourth agent. Preoperatively, these high-risk patients received cisplatin either intraarterially or intravenously. RESULTS: 171 eligible patients were entered, of which 128 were stratified into the high-risk group. When all 171 were analysed by intention-to-treat, actuarial overall and event-free survival rates at ten years were 72% and 66%, respectively. No benefit of intraarterial cisplatin application was detected. Cumulative treatment toxicity was considerable. CONCLUSIONS: In a multicenter setting, intensive treatment of osteosarcoma according to protocol COSS-86 led to long-term disease-free survival for two thirds of patients. We saw no benefit of using the intraarterial route to administer cisplatin.     

Modified Bagolini striated glass test: clinical applications of starlight test in binocular visual field screening

AIM: To introduce the "starlight" test which was devised to check binocular vision in normal conditions of seeing in a rapid, easy, and cost effective manner and to estimate the possibility of its clinical use in screening the binocular visual field of patients. METHOD: The Bagolini striated glass test consists of optically plano lenses with imperceptible parallel scratches that barely blur the environment but produce two perpendicular luminous stripes (right eye stripe of 45 degrees and left eye stripe of 135 degrees) when subjects with normal binocular vision view one light source. Unlike the original Bagolini test, the starlight test uses three light sources in horizontal or vertical lines according to the testing purposes and the subject is asked to fixate upon the centre light. Through Bagolini glasses, the subject observes the resulting grid-like pattern and the state of binocular visual field of the subject can be roughly estimated. RESULTS: Normal subjects and patients with strabismus, visual field loss from intracranial diseases, glaucoma, retinitis pigmentosa, and functional visual loss were examined using the starlight test and findings from each case were discussed. CONCLUSIONS: The starlight test, which was made by hand at a low cost, is a simple test that can be used clinically. It provides information about the state of binocular vision of patients in normal conditions of seeing. It is also useful because it enables the examiner to share similar experiences with the examinee. The results suggest it can be effective in visual field screening.     

Maturation of rat dendritic cells during intrahepatic translocation evaluated using monoclonal antibodies and electron microscopy

Specific populations of hepatic sinusoidal cells were stained with monoclonal antibodies that recognize monocytes/macrophages (ED1), tissue macrophages (Kupffer cells) (ED2), MHC class II (Ia) antigen (MRC OX6), and dendritic cells/gamma,delta T-cells (MRC OX62) and analyzed by light and electron microscopy. The majority of ED1(+) and/or ED2(+) cells were localized to the hepatic parenchyma, whereas OX6(+) and/or OX62(+) cells were more densely distributed within Glisson's sheath than in the hepatic parenchyma. Double-immunoperoxidase staining of normal liver for ED1, ED2, and OX6 identified dendritic cells (DC) of two different phenotypes, ED1(+)ED2(-)OX6(+) and ED1(-)ED2(-)OX6(+). DC can be classified into three different types based on ultrastructural characteristics. The first type (type I) is characterized by one or more long cytoplasmic processes and a well-developed lysosomal system. The second type (type II) has an inconspicuous lysosomal system, abundant hyaloplasm, and characteristic short cytoplasmic processes. The third type (type I-II) has cytologic features intermediate between those of type I and type II DC. At the electron-microscopic level, these three cell types are found in the sinusoidal lumen, whereas the majority of type II DC are located in the space of Disse and Glisson's sheath. Furthermore, some OX6-labeled elongated DC appeared to traverse the lumen of sinusoids through endothelial pores to enter the space of Disse. One hour after intravenous injection of latex particles (0.81 micrometer in diameter), numerous latex-laden dendritic cells (ED1(+)OX6(+), type I and type I-II) were detected in the lumen of hepatic sinusoids, but not in the space of Disse or Glisson's sheath. These findings suggest that normal rat liver contains resident dendritic cells which downregulate phagocytic activity and mature into potent accessory cells during migration from the portal vein toward the central vein. These DC then traverse the sinusoidal lumen to the hepatic lymph system via the space of Disse.     

Efficacy of exogenous fibronectin in wound healing in malnourished rats

BACKGROUND/PURPOSE: Fibronectin (FN) plays an important role in fibrin matrix formation during the wound healing process. The authors investigated whether exogenous FN increases the bursting strength (BS) of surgical wounds in malnourished rats. METHODS: Ninety rats were grouped according to three nutritional conditions (n = 30 in each group). All animals underwent a transverse celiotomy after 3 weeks of feeding, and FN (2 mg/body/day) was given postoperatively to 15 rats in each of the three groups. Wound BS was measured on postoperative days (PODs) 3, 5, and 7 after removal of the sutures. RESULTS: BS in normally nourished rats (group N; 142.6+/-23.4 mmHg) was significantly higher than that in protein malnourished rats (group PM, 110.2+/-11.2 mm Hg) and protein/ calorie (Cal)-malnourished rats (group PCM, 76.5+/-10.7 mm Hg) on POD 7 (P<.01). However, BS values for groups PM + FN (147.0+/-21.1 mmHg) and PCM + FN (115.1+/-28.9 mm Hg) were intensified significantly in comparison with groups PM and PCM (P<.01). Plasma FN levels in rats of the three FN nontreated groups were similarly decreased on POD 3 or 5, but returned to the preoperative level on POD 7, whereas those for the other three FN-treated groups increased after POD 3. CONCLUSION: Intravenous administration of FN might strengthen the weakened wounds of malnourished animals.     

The relation between platelet aggregation and constitutional and lifestyle variables in two Japanese communities

To investigate the contribution of the platelet aggregation in the development of cardiovascular diseases, we examined the relation of constitutional and lifestyle variables with platelet aggregation for a total of 306 males aged 50 to 70 in Ikawa town, Akita prefecture (n = 163) and Noichi town, Kochi prefecture (n = 143). The examination of platelet aggregation was completed within 3 hours of obtaining blood samples. We used ADP (Adenosine 5'-diphosphate) as an agonist and obtained PATI (the platelet aggregatory threshold index) by nephelometry. Platelet count, mean platelet volume, white blood cell count, serum fatty acid compositions were also examined and dietary intake of fish, seafood and soy bean foods were inquired using one-week dietary records. PATI indicated a logarithmic normal distribution in both Ikawa and Noichi. The mean of logarithmic transformed PATI (log PATI) was higher in Ikawa than in Noichi. Thus platelet aggregation was lower in Ikawa than in Noichi. According to multiple regression analysis, age, platelet count in platelet rich plasma, mean platelet volume in platelet rich plasma, and white blood cell count were inversely associated with log PATI. Serum arachidonic acid composition tended to be inversely related with log PATI. Serum n3-polyunsaturated fatty acid composition was positively related with log PATI, and log gamma-GTP tended to be positively associated with log PATI. Soy protein intake and cigarette smoking showed no consistent associations with log PATI. This cross-sectional study suggests that serum n3-polyunsaturated fatty acid, and gamma-GTP, as an index of alcohol intake, reduce platelet aggregation while age, white blood cell count, platelet count, mean platelet volume, and serum arachidonic acid raise platelet aggregation.