Results of animal experiments using an undulation pump total artificial heart: analysis of 10 day and 19 day survival

An undulation pump is a special rotary blood pump in which rotation of a brushless DC motor is transformed to an undulating motion by a disc in the pump housing attached by means of a special link mechanism. In the blood pump, a closed line between the disc and housing moves from the inlet to the outlet by this undulating disc motion, which sucks and pushes the blood from the inlet to the outlet. Because the same phenomena occurs at both sides of the disc, a continuous flow is obtained when the motor rotational speed is constant. The pump flow pattern can be easily changed from continuous flow to pulsatile flow by controlling the motor drive current pattern. A seal membrane made of segmented polyurethane protects the blood from invading the link mechanism as well as the motor. UPTAH is fabricated with two undulation pumps and two brushless DC motors. Its size is 75 mm in diameter and 80 mm long, and it has one of the great advantage of no compliance chamber required in the system. UPTAHs were implanted under cardiopulmonary bypass (CPB) into the chest cavities of 16 goats, each weighing between 41 and 72 kg. No anticoagulant and antiplatelet agent was used after the surgery. The left atrial pressure was automatically controlled to prevent its elevation and sucking of the atrial wall into the atrial cuff. The following results were obtained: (1) UPTAHs fit well into all the goats; (2) the longest survival was 19.8 days, the cause of death was bleeding from the aortic anastomosis; (3) No thrombus was observed in the blood pump despite no anticoagulant use. Hemolysis depended upon the length of CPB during surgery. When CPB time was within 2 hours, hemolysis level returned to baseline within a few days of the surgery. UPTAH is a promising implantable TAH, because of its small size and easy controllability.     

Case report of de novo dup(18p)/del(18q) and r(18) mosaicism

This is a report of a 27-year-old woman with an unusual de novo chromosomal abnormality. Mosaicism was identified in peripheral blood cells examined by standard G-bands by trypsin using Giemsa (GTG) analysis and fluorescence in situ hybridization (FISH) analysis with chromosome-18 region-specific probes, 46,XX,del(18)(pter → q21.33:)[41], 46,XX,r(18)(::p11.21 → q21.33::)[8], and 46,XX,der(18)(pter → q21.33::p11.21 → pter)[1]. On the other hand, the karyotype of periodontal ligament fibroblasts was nonmosaic, 46,XX, der(18)(pter → q21.33::p11.21 → pter)[50]. All cell lines appeared to be missing a portion of 18q (q21.33 → qter). The pattern of the dup(18p)/del(18q) in the rod configuration raises the possibility of an inversion in chromosome 18 in one of the parents. However, no chromosomal anomaly was detected in either parent. The most probable explanation is that de novo rod and ring configurations arose simultaneously from an intrachromosomal exchange. The unique phenotype of this patient, which included primary hypothyroidism and primary hypogonadism, is discussed in relation to her karyotype.     

Anti-CD45R antibody-treated normal lymphocytes respond to interleukin 2 (IL2) after stimulation by allergens

Interleukin 2 (IL2) responsiveness was specifically induced by Dermatophagoides farinae (Df) antigen in Df-sensitized lymphocytes from asthmatic children, but not in normal lymphocytes. Df-induced IL2 responsiveness was also observed in normal lymphocytes pretreated (Day 0) with anti-CD45R antibody, which recognize suppressor inducer subset among CD4+ T cells. However anti-CD45R antibody was no longer effective when the lymphocytes were cultured for more than one day with the antigen, suggesting its effect in the initial phase of the reaction. The intensity of the response induced in normal lymphocytes by the anti-CD45R was comparable to that of the patients sensitized to the nominal antigen. The response of the patients was no longer augmented by the anti-CD45R antibody. Taken together, these data suggest that even normal lymphocytes have potentiality to elicit Df-induced IL2 responsiveness and it is probably derepressed by inhibiting suppressor inducer subset with the anti-CD45R antibody. Also suggested is a defective suppressor inducer activity in the lymphocytes which may lead to hyperreactivity to allergens in asthmatic children.     

Fundamental study on the mechanism of DNA degradation in tissues fixed in formaldehyde.

The mechanism of DNA degradation and its clinical applications were examined. When purified lambda phage and extracted liver DNA were fixed in phosphate buffered formaldehyde, the DNA did not degrade, but there was incomplete digestion with endonuclease. Rat liver tissues were fixed under various conditions and DNA extracted. Immediate fixation with buffered formaldehyde at low temperature, or the addition of EDTA to buffered formaldehyde blocked the DNA degradation. Analysis of pulsed field gel electrophoresis also showed that DNA was degraded before extraction. These results suggest that tissue nuclease has an important role in DNA degradation in tissue. Furthermore, formaldehyde fixation at low temperature, which may take time and which decreases slightly the staining capacity, is useful for the extraction of intact DNA. For clinical application, the detection of provirus was examined. Genomic DNA was extracted from a necropsy sample of adult T cell leukaemia fixed in formaldehyde; human T cell leukaemia virus type-I (HTLV-I) provirus was successfully detected by Southern blotting.     

Role of the spleen in immunosuppression of gastric cancer: predominance of suppressor precursor and suppressor inducer T cells in the recirculating spleen cells.

In order to analyse the role of the spleen on immunosuppression of gastric cancer, T cell phenotypes in the spleen cells (SC) were investigated by two colour fluorescence flow cytometry, with reference to their suppressor cell activity. Suppressor T cell phenotypes of CD4+2H4+ cells (suppressor/inducer T cells) and CD8+CD11+ (suppressor T cells) were distributed predominantly in SCs from patients with gastric cancer, while they were distributed scarcely in those with liver cirrhosis. Moreover, CD4+2H4+ cells and CD8+CD11+ cells were found predominantly in SCs and splenic vein lymphocytes (SVL) respectively. Among SCs, a significantly higher proportion of CD4+2H4+ cells was found in the recirculating SCs, but fewer were found in the residual SCs. Higher activity of Concanavalin-A induced suppressor cells was found in the former and that of spontaneously activated suppressor cells was found in the latter. These results suggest the suppressor precursor and suppressor/inducer T cells might distribute predominantly in the cells recirculating from the spleen, and that suppressor cells might be matured during the migration from the spleen.     

Monocyte activating factor in sarcoidosis. I. Existence of the factor in sarcoidosis sera

Sarcoidosis sera were found to have the ability to induce normal human monocytes to spread. Gel filtration of sarcoidosis sera on Sephadex G-200 showed that the factor mainly responsible for this activity had a molecular weight of about 70,000. The spreading factor also possessed the ability to increase all cell size of normal human monocytes as well as to increase their phagocytosis and glucose consumption. Accordingly, the spreading factor seems to be considered as a monocyte activating factor. Sarcoidosis sera showed a macrophage migration inhibitory activity, as well. On Sephadex G-200 column chromatography of the sera, the most obvious inhibitory activity was eluted in the fraction with a molecular weight of about 45,000. The macrophage migration inhibitory factor had the ability neither to increase cell size of normal human monocytes nor to increase their phagocytosis and glucose consumption.     

Three-dimensional two-layer collagen matrix gel culture model for evaluating complex biological functions of monocyte-derived dendritic cells

Dendritic cell-like cells (Mo-DCs) generated from peripheral blood monocytes with interleukin-4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF) have been used as tools to treat cancer patients (DC-vaccines). Because Mo-DCs have multiple antigen presentation-related functions, including phagocytosis, migration, cytokine production, and T cell stimulation, establishment of a method for simultaneously evaluating the various functions of Mo-DCs is important. We developed a new in vitro three-dimensional two-layer collagen matrix culture model that consists of a collagen gel containing Mo-DCs as the lower layer and a collagen gel containing necrotic GCTM-1 tumor cells and/or T cells as the upper layer. We used this system to observe simultaneously multiple functions of Mo-DCs by phase-contrast or fluorescence microscopy and to assess IL-12 secretion during more than 2 weeks of culture. We also observed interactions between Mo-DCs and necrotic GCTM-1 or T cells on an individual cell basis by time-lapse videomicroscopy. In addition, we collected Mo-DCs from the collagen gels by collagenase treatment and analyzed the expression of antigen presentation-related molecules such as HLA-DR, CD80, CD83, and CD86 on Mo-DCs. This model may be a useful tool for evaluation of the various functions of Mo-DCs used as DC vaccines and for studies of the complex behaviors of Mo-DCs in vivo.     

Harderian gland dependency of immunoglobulin A production in the lacrimal fluid of chicken

Involvement of the Harderian gland (HG) in the production of lacrimal immunoglobulin (especially IgA) was investigated. The lacrimal concentration of each immunoglobulin class was not affected by surgical bursectomy but was reduced by cyclophosphamide (CY) and testosterone (TP) treatments. Surgical removal of the Harderian gland caused a remarkable reduction of both the lacrimal concentration of each immunoglobulin class and the specific antibody titre, and and IgA was almost undetectable. The lacrimal concentration of each immunoglobulin class, as well as the specific antibody titre, was not affected by surgical removal of the Lacrimal gland (LG). The route of antigen administration produced no difference in the class of lacrimal immunoglobulin produced. The results indicate that the production of immunoglobulin in chicken tears may be dependent on the HG and that lacrimal immunoglobulin may be synthesized and secreted locally in the HG. Lymphocytes of the HG are of bursa of Fabricius origin and are seeded into the HG prior to hatching and its lymphocytes do not appear to be involved in systemic immunity.     

Rat strain differences in the early stage of porcine-serum-induced hepatic fibrosis.

Rat strain differences in the early development of porcine serum (PS)-induced hepatic fibrosis were histologically and immunohistochemically examined using Brown Norway (BN), Sprague Dawley (SD) and Wistar rats. They were injected i.p. with 0.5 ml sterile PS twice a week for 4 and 8 weeks. In addition, rats treated with physiological saline in the same way served as controls. At 4 weeks, hepatic fibrosis accompanying fibrous septa mainly composed of type III collagens developed in BN and SD rats but not in Wistar rats. In addition, the numbers of eosinophils, CD3-positive cells and ED-1-positive cells significantly increased in BN and SD rats, that of CD45RA-positive cells in BN rats, and that of alpha-smooth muscle actin (SMA)-positive cells in SD rats, respectively. Such differences in the number of inflammatory cells may be related with the absence of hepatic fibrosis in Wistar rats at 4 weeks. At 8 weeks, hepatic fibrosis with formation of many small-sized pseudolobules was observed in all strains at almost similar degree, and the numbers of infiltrating cells increased in all strains of rats with some exception. In addition, the main location of inflammatory cells was different, suggesting a different role of each inflammatory cell in the process of hepatic fibrosis.