Estimation of generator potentials in primary spindle endings evoked by brief tendon taps in man

The ligamentum patellae in human subjects was tapped by a random triangular waveform, while single primary spindle afferent spikes were recorded from the femoral nerve. Cross-correlation between the primary spindle afferent spikes and the taps revealed a prominent peaked distribution of the discharge probability. The cross-correlograms showed that response time from the onsets of taps to the afferent spike was 7.5 +/- 0.6 msec in 6 units and that the width between the feet of the peaked distribution was 3.3 +/- 0.2 msec in 6 units. The width of the peak implies jitter of the response time of the primary spindle spikes which appear to correspond with the time-to-peak of generator potentials elicited by the taps in primary endings of muscle spindles. Integration of the peaked distribution showed a slow slope at the beginning and near the summit and a steep slope on the way. This curve is assumed to represent the rising phase of the generator potentials in primary spindle endings.     

Change of Mouse CD5+ B1 Cells to a Macrophage-Like Morphology Induced by Gamma Interferon and Inhibited by Interleukin-4

The in vitro effects of gamma interferon (IFN-γ) on the mouse CD5⁺ B1-cell line, TH2.52, a hybridoma between mouse B lymphoma and mouse splenic B cells that expresses a series of B1 markers, were investigated. A significant number of macrophage-like cells appeared in the cultures of TH2.52 cells exposed to IFN-γ, these adhering to plastic dishes and exhibiting phagocytic activity. Positive for esterase staining, the macrophage-like cells returned to the original TH2.52 morphology upon removal of IFN-γ. The change was prevented by treatment with SB202190, an inhibitor of p38 mitogen-activated protein (MAP) kinase and by transfection of a p38 MAP kinase dominant-negative mutant. Further, interleukin-4 (IL-4) inhibited IFN-γ-induced phosphorylation of p38 MAP kinase and the appearance of macrophage-like cells. IFN-γ and IL-4 exhibited contradictory actions on morphological change of CD5⁺ B1 cells into macrophage-like cells. Differential regulation of CD5⁺ B1 cells by IFN-γ, a Th1 cytokine, and IL-4, a Th2 cytokine, may have clear immunological significance.     

Augmentation of nitric oxide production by gamma interferon in a mouse vascular endothelial cell line and its modulation by tumor necrosis factor alpha and lipopolysaccharide

The effect of gamma interferon (IFN-gamma), tumor necrosis factor alpha (TNF-alpha), and lipopolysaccharide (LPS) on nitric oxide (NO) production in the mouse vascular aortic endothelial cell line END-D was examined. LPS, TNF-alpha, and a low concentration of IFN-gamma inhibited NO production in END-D cells, while a high concentration of IFN-gamma definitely enhanced it. The NO production induced by a high concentration of IFN-gamma was further augmented by using IFN-gamma in combination with LPS or TNF-alpha. In sequential incubations of LPS and IFN-gamma, the enhancement of NO production required prior treatment with IFN-gamma. Stimulation of END-D cells with a high concentration of IFN-gamma led to the expression of inducible NO synthase (iNOS). The augmentation of NO production by IFN-gamma alone or in combination with LPS or TNF-alpha was completely blocked by several inhibitors of iNOS. It was strongly suggested that a high concentration of IFN-gamma itself enhanced NO production in END-D cells through inducing the expression of iNOS. LPS and TNF-alpha exclusively modulated the activity of iNOS once its expression was triggered by IFN-gamma. On the other hand, a low concentration of IFN-gamma, LPS, and TNF-alpha reduced NO production through down-regulating constitutive NOS (cNOS). The differential regulation of cNOS- and iNOS-mediated NO production by IFN-gamma, TNF-alpha, and LPS is discussed.     

Inhibition of p38 mitogen-activated protein kinase augments lipopolysaccharide-induced cell proliferation in CD14-expressing Chinese hamster ovary cells

CD14-expressing Chinese hamster ovary (CD14-CHO) cells, established by transfection of human CD14 DNA, acquired high responsiveness to lipopolysaccharide (LPS) through membrane-bound CD14 expression. LPS induced DNA synthesis and activated a series of mitogen-activated protein (MAP) kinases, extracellular signal-regulated kinase 1/2 (Erk1/2), p38, and c-Jun N-terminal kinase/stress-activated protein kinase, in CD14-CHO cells but not in mock-transfected CHO cells. Anti-CD14 antibody completely abrogated both LPS-induced DNA synthesis and LPS-induced phosphorylation of those MAP kinases, suggesting a critical role of membrane-bound CD14 in LPS signaling. A p38 MAP kinase inhibitor, SB203580, markedly augmented LPS-induced DNA synthesis in CD14-CHO cells, whereas an Erk1/2 inhibitor, PD98059, had no affect. On the other hand, SB203580 exhibited no effect on epidermal growth factor-induced DNA synthesis in CD14-CHO cells, although PD98059 inhibited it significantly. The activation and inactivation of p38 MAP kinase with dominant negative and dominant positive mutants also suggested the participation of p38 MAP kinase in LPS-induced DNA synthesis. It was therefore suggested that the activation of p38 MAP kinase can negatively regulate LPS-induced cell proliferation in CD14-CHO cells.     

Marked reduction of mouse peritoneal CD5+ B cells by intraperitoneal administration of lipopolysaccharide.

Intraperitoneal administration of lipopolysaccharide to mice induced a marked reduction of CD5+ B cells in the peritoneal cavity. The reduction was not induced by intravenous, subcutaneous, or oral administration of lipopolysaccharide. The reduction continued for about 10 days after the injection, and the CD5+ B-cell count recovered to the normal state about 14 days after the injection. The reduction of peritoneal CD5+ B cells might be caused by apoptotic cell death. Injection of lipopolysaccharide did not result in production of antibody to lipopolysaccharide. On the other hand, intraperitoneal injection of heat-killed bacteria did not induce a reduction of peritoneal CD5+ B cells and elicited the definite production of antibody to lipopolysaccharide.     

Requirement of the IFN-alpha/beta-induced CXCR3 chemokine signalling for CD8+ T cell activation

BACKGROUND: Activation of both CD4+ T and CD8+ T cells is triggered by the engagement of the T cell antigen receptor (TCR) with MHC/peptide complexes on antigen-presenting cells. This process also requires other molecular interactions, which transmit co-stimulatory signals to these T cells. To ensure an effective immune response, distinct T cell subsets may additionally employ unique mechanism(s) for efficient activation. RESULTS: We here show that mutant CD8+ T cells lacking the IFN-alpha/beta signalling components are hyporesponsive to antigen stimulation in vitro. We further show that IFN-alpha/beta-mediated signals are required for induction of the chemokines IP-10/I-TAC and their common receptor, CXCR3, and in turn provide evidence that CXCR3-mediated signals indeed function in the activation and proliferation of CD8+ T cells, particularly for the CD44low naive phenotype cells. CONCLUSION: The CXCR3 chemokine system is regulated by IFN-alpha/beta in CD8+ T cells, and it is critical for the efficient cell activation. The present study therefore reveals a novel role of the IFN-alpha/beta-CXCR3 signalling cascade in CD8+ T cell activation.     

Left ventriculotomy of the heart: tissue repair and localization of collagen types I,II, III,IV, V,VI and fibronectin

Summary The reparative process following left ventriculotomy was investigated immunohistochemically using anti-type I, II, III, IV, V and VI collagen antibodies, and anti-fibronectin antibody. Wound healing began with proliferation of young fibroblasts positive for type I, III and V collagens at the wound margin; vascular granulation tissue then grew into the injured myocardium followed by deposition of fibrous components immunoreactive with type I and III. At 30 days after operation when almost the entire thickness of the myocardium at the wound was replaced by fibrosing granulation tissue, a small cluster of adipocytes appeared around capillaries at the wound margin. The granulation tissue was gradually replaced by the adipose tissue with establishment of a fibrous union at the subendocardium by 90 days. In addition to type I and III collagens, type VI collagen was detected in a fine fibrillary pattern along thick collagen fibre bundles in the fibrous tissue and around the adipocytes. Fibronectin was distributed diffusely in the granulation tissue and gradually decreased with increase of the fibrous components. These results indicate that the ventriculotomy was finally repaired in the form of a fibrous scar, particularly in the endocardium. There was marked infiltration of adipose tissue in the damaged myocardium. Presumably type VI collagen, as well as type I and type III collagens, plays an important role in wound union.     

Detection of antibodies to the Epstein-Barr virus nuclear antigens in the sera from patients with systemic lupus erythematosus

The sera from 65 patients with systemic lupus erythematosus (SLE) were examined by the immunoblotting method to detect antibodies to Epstein-Barr virus (EBV)-associated antigens, especially EBV nuclear antigens (EBNA), and compared with the sera from 66 healthy subjects roughly age- and sex-matched to the patients. Most sera from patients with SLE defined three major EBV-associated antigens with molecular weights (MW) of 70,000 (70K), 90K and 140K in Raji cells, which must correspond to the EBNA-1, 2, and 3, respectively. Approximately 70% of the sera from SLE patients demonstrated the antibodies to the 90K and 140K antigens, whereas the positive rates of these two antibodies were less than 10% in the sera from healthy subjects. The differences of these positive rates of the antibodies between SLE patients and healthy subjects were statistically highly significant. Antibody to EBNA-1 was conspicuously detected in the sera from both SLE patients and healthy subjects, although the difference between the two groups was still significant. The possible role of EBV infection was discussed on the basis of the pathogenesis of SLE.     

Photocatalytic hydrogenation of nitrobenzene to aniline over titanium(iv) oxide using various saccharides instead of hydrogen gas

Bare TiO₂ photocatalyst almost quantitatively converted nitrobenzene to aniline with various saccharides without the use of hydrogen gas. Although aniline was formed when any saccharide was used, the use of disaccharides (lactose, maltose, and sucrose) decreased the reaction rate. The rate of photocatalytic hydrogenation of nitrobenzene using saccharides is determined by the degradation rate of saccharides at positive holes. When glucose was used, formic acid, arabinose, glyceraldehyde and lactic acid were obtained, which are products that are consistent with the product of the photocatalytic oxidation of glucose.

10 kinds of saccharides were investigated as hydrogen source in photocatalytic hydrogenation of nitrobenzene to aniline.     

Differential expression of autophagy in Hodgkin lymphoma cells treated with various anti-cancer drugs

AIM: autophagy is a pivotal physiological process for survival during starvation, differentiation and normal growth control. It is defined as the process of sequestrating cytoplasmic proteins or even entire organelles into the lytic compartment (lysosome/vacuole). This study investigates the expression of autophagy in Hodgkin lymphoma cells treated with various anti-cancer drugs. METHODS: Hodgkin's lymphoma cells (HD-My-Z cells) were cultured with various anti-cancer drugs, such as bleomycin, adriamycin, gemcitabine and paclitaxel. Autophagy was detected by fluorescent pattern of light chain 3(LC3) proteins and the apoptotic cell death was determined by annexin V binding. RESULTS: autophagy was detected in HD-My-Z cells treated with gemcitabine, but not with bleomycin, adriamycin and paclitaxel. Adriamycin exhibited the strongest cytotoxic action, and the cytotoxic action of bleomycin and gemcitabine was less marked compared with adriamycin. Paclitaxel did not cause significant cell death in the cells. CONCLUSION: autophagy was differentially expressed in Hodgkin lymphoma cells treated with anti-cancer drugs and the expression did not correspond to the apoptotic cell death.