Rheological properties, oxidative stability, and tocopherol content during storage of fried dough made with Silky fowl egg: comparison with hen egg.

Eggs from Silky fowl and White Leghorn hens were used to prepare fried dough. The rheological properties, lipid oxidative stability, and trans, trans-2,4-decadienal and tocopherol content of fried dough made with Silky fowl egg were compared with dough made with hen egg. The fried dough was stored in a glass bottle at 50 degrees C in the dark for 12 d. The fried dough made with Silky fowl egg showed little change in hardness and adhesion for 12 d at 50 degrees C. However, in the fried dough made with hen egg, hardness increased drastically and adhesion decreased. The fried dough made with Silky fowl egg showed restricted generation of hydroperoxides during 12 d in storage at 50 degrees C. In contrast, the fried dough made with hen egg showed an increased amount of hydroperoxides during the 12-d storage. The lowest concentration of trans, trans-2,4-decadienal was observed in fried dough made with Silky fowl egg, whereas the concentration of trans, trans-2,4-decadienal in fried dough made with hen egg was significantly increased. Total tocopherols in fried dough made with Silky fowl egg were degraded 23.3 mg/100 g of fried dough by the end of the experimental period at 50 degrees C. In contrast, total tocopherols in the fried dough made with hen egg were degraded 40 mg/100 g of fried dough. The ratio of unsaturated fatty acids to saturated fatty acids decreased and the hydroperoxide content increased with storage time. The unsaturated fatty acid:saturated fatty acid ratio and hydroperoxide and tocopherol contents were lower in fried dough made with Silky fowl egg than in that made with hen egg, indicating decreased lipid oxidation. The present experiment suggests that the use of Silky fowl egg could improve the rheological properties, oxidative stability, and trans, trans-2,4-decadienal and tocopherol contents of fried dough.

Nurse-like cells from bone marrow and synovium of patients with rheumatoid arthritis promote survival and enhance function of human B cells.

Thymic nurse cells are known to interact with T cells and play a role in their functional maturation. However, the role of nurse cells in B cell maturation and differentiation is less well established, especially at extralymphoid sites. To address this issue, nurse-like cell clones from bone marrow and synovial tissue of patients with RA (RA-NLC) were established and characterized. RA-NLC constitutively expressed CD29, CD49c, CD54 (ICAM-1), CD106 (VCAM-1), CD157 (BST-1), and class I MHC molecules, and secreted IL-6, IL-7, IL-8, granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF). Bone marrow-derived and synovial RA-NLC differed in that the former secreted IL-7 and expressed a greater density of CD157 constitutively and after stimulation with IFNgamma, whereas the latter secreted G-CSF and more IL-6. Stimulation of both bone marrow and synovial RA-NLC induced expression of CD40 and class II MHC, but not CD154 (CD40L) or CD35. RA-NLC rescued peripheral B cells from spontaneous apoptosis and promoted survival of B cells for > 4 wk. B cell survival was blocked by antibodies to CD106 or CD157. RA-NLC also increased Ig production from B cells. After long-term culture (4-6 wk) with RA-NLC, but not alone or with fibroblasts, outgrowth of B cells was observed. All B cell lines derived from these cultures had been transformed by EBV, although the RA-NLC themselves were not infected with EBV. Precursor frequency analysis indicated that approximately 1 in 12,500 peripheral B cells could give rise to these EBV-transformed B cell lines upon coculture with RA-NLC. These results indicate that RA-NLC from bone marrow and synovium have the capacity to rescue B cells from spontaneous apoptosis, facilitate Ig production, and promote the outgrowth of EBV-transformed B lymphoblastoid cells. These findings suggest that RA-NLC may play a role in the local and systemic hyperreactivity of B cells characteristic of rheumatoid arthritis.

Quantitative analysis of the usage of human T cell receptor alpha and beta chain variable regions by reverse dot blot hybridization.

We previously developed an adaptor ligation-mediated PCR method to amplify the T cell receptor (TCR) cDNA pools. In the present study we applied reverse dot blot hybridization to PCR-amplified specimens for quantitative analysis of the usage of TCR alpha and beta chain variable (V) region. 44 VA sequence-specific oligonucleotide probes (SSOPs) and 38 VB SSOPs were synthesized corresponding to unique sequences of VA and VB subfamilies. Peripheral blood lymphocytes of ten healthy donors and five T cell clones established from bone marrow cells were examined for VA and VB usage using this method. The results were consistent with those obtained by a colony hybridization method and those by immunofluorescence staining using monoclonal antibodies to VA and VB. Thus, reverse dot blot hybridization for TCR V(alpha) and Vbeta is a new, easy and dependable technique useful for analysis of VA and VB usage by human T cells.

Proinflammatory cytokines detectable in synovial fluids from patients with temporomandibular disorders.

OBJECTIVE: To measure the levels of the proinflammatory cytokines, interleukin (IL)-1 beta, IL-6, tumor necrosis factor- (TNF) alpha, IL-8, and interferon- (IFN) gamma in synovial fluid samples taken from patients with temporomandibular disorders (TMD). STUDY DESIGN: We studied 6 asymptomatic volunteers and 51 patients with TMD. The IL-1 beta, IL-6, TNF-alpha, IL-8, and IFN-gamma levels in temporomandibular joint synovial fluid were measured using enzyme-linked immunosorbent assay. RESULTS: Measurable level of at least one cytokine in the synovial fluid was found in 40 (64.5%) of 62 joints in the patients: IL-1 beta and IFN-gamma were each detected in 18 (29.0%) of 62 joints; IL-6 in 13 (21.0%) of 62 joints; IL-8 in 11 (19.3%) of 57 joints; and TNF-alpha in only 5 (8.1%) of 62 joints. None of these cytokines was detectable in the synovial fluid in the control group. Furthermore, there was a strong correlation between the detection of IL-1 beta and pain in the joint area. CONCLUSIONS: These data clearly demonstrate increased levels of several proinflammatory cytokines in certain patients with TMD and suggest that these cytokines may play a role in the pathogenesis of synovitis and degenerative changes of the cartilaginous tissue and bone of the temporomandibular joint.

Study of wall effect on the flow of milk in capillary.

Dependence of the viscosity of milks on the bore-size of capillary viscometer has been studied at 25 degrees C. Viscosity measurements were carried out with a Maron-Belner type low shear capillary viscometer with various capillary radii (0.0190-0.076 cm) within the shear stress range of 0.2-30 dynes/cm2 at the capillary wall. Each milk showed non-Newtonian flow behavior and dependence on the size of the capillary. The viscosity of human fresh milk and cow's fresh milk decreased with the decreases in the capillary bore-size within the range of shear stress studied, and the viscosity of homogenized milk, although similar phenomena as above were observed in higher shear stress, decreased with the decrease in the capillary bore-size at the range of lower shear stress. The difference in characteristics of capillary bore-size dependence on viscosity may be attributed to the difference in the behavior of fat droplets existing near the wall at which thermal motion of fat droplets will be hindered by the existence of capillary wall. The results were represented by the modified Ree-Eyring generalized flow formula for a flow system containing one Newtonian flow unit and one non-Newtonian flow unit which included additional terms dependent on the capillary bore-size. The viscosity equation suitable for analysing the dependence of viscosity on the capillary bore-size at entire ranges of rate of shear, was derived by assuming the wall layer. Composed of double layers (of thickness do and di - do) in the flowing liquid in capillary tube, and by combining this equation with the modified Ree-Eyring equation, the values of do and di were calculated.

Monoclonal antibodies to feline lymphocyte membranes recognize the leukocyte-common antigen (CD45R).

By immunization of BALB/c mice with a feline T lymphoblastoid cell line, MYA-1 cells, two types of lymphocyte-specific monoclonal antibodies (mAbs) were obtained. The 220/205/190 kd protein defined by 2F11 mAb is highly expressed on the surface of MYA-1 cells and another feline T lymphoma cell line, FL74 cells. The protein is also expressed on normal feline thymocytes, splenocytes and feline peripheral blood mononuclear cells (PBMCs). Another mAb, 17B10, caused similar results as those of 2F11 except for its low reactivity with FL74 cells. The second type of mAb, 15B3, defined the 220 kd protein. The reactivities of this mAb with MYA-1 cells, FL74 cells, PBMCs and feline splenocytes were lower than the former two mAbs, and did not react to feline thymocytes. On the other hand, 17B10 and 15B3 defined partial populations of MYA-1 and FL74 cells recognized by 2F11. The cells defined by the 2F11 and 17B10 are all leukocytes in spleen and lymph node. In contrast, 15B3 defined most of the cells in B cell area and partially in T cell area. These results suggested that 2F11 and 17B10 recognized the specific antigen of 220/205/190 kd of the leukocyte-common antigen (L-CA) family, CD45R, with different epitopes, and that 15B3 defined the distinct antigen of 220 kd on CD45R.

Eight-year observation and comparative study of specific pathogen-free cats experimentally infected with feline immunodeficiency virus (FIV) subtypes A and B: terminal acquired immunodeficiency syndrome in a cat infected with FIV petaluma strain.

Three specific pathogen-free cats experimentally infected with feline immunodeficiency virus (FIV) strains Petaluma, TM1 and TM2, respectively were observed for over 8 years. Without showing any significant clinical signs of immunodeficiency syndrome (AIDS) for 8 years and 4 months of asymptomatic phase, the Petaluma-infected cat exhibited severe stomatitis/gingivitis, anorexia, emaciation, hematological and immunological disorders such as severe anemia, lymphopenia, thrombocytopenia, and decrease of CD4/CD8 ratio to 0.075, and finally died with hemoperitoneum at 8 years and 8 months post-infection. Histopathological studies revealed that the cat had systemic lymphoid atrophy and bone marrow disorders indicating acute myelocytic leukemia (aleukemic type). Plasma viral titer of the cat at AIDS phase was considerably high and anti-FIV antibody titer was slightly low as compared with the other FIV-infected cats. In addition, immunoblotting analysis using serially collected serum/plasma samples of these cats revealed that antibodies against FIV proteins were induced in all the infected cats, however in the Petaluma-infected cat anti-Gag antibodies disappeared during the asymptomatic period. These results suggested that plasma viral load and anti-FIV Gag antibody response correlated with disease progression, and supported FIV-infected cats as a suitable animal model of human AIDS.

Antioxidant effects of the water-soluble fraction of baked sponge cake made with silky fowl egg: comparison with White Leghorn egg.

1. The antioxidant effects of the water-soluble fraction of baked sponge cakes made with silky fowl eggs and White Leghorn eggs were studied. The mechanism of the antioxidant effect was also investigated. 2. The antioxidant effect on the oxidation of linoleic acid increased in the water-soluble fraction of cake made with silky eggs. In contrast, Leghorn eggs significantly decreased the rate of antioxidant activity. The browning index of the water-soluble fraction of baked sponge cake made with silky fowl eggs changed from 0.052 to 1.240 after 20 min at 180 degrees C, while that made with Leghorn eggs changed from 0.037 to 0.710. 3. There are correlations between the rate of browning index and antioxidant activity. Superoxide anion (O2(-)) and hydrogen peroxide (H(2)O(2)) in water-soluble fractions of baked sponge cakes made with silky fowl eggs and hen's eggs were formed during light exposure for 20 min at 10,000 lux, and their formation could be significantly inhibited by the addition of tryptophan or mannitol, scavengers of hydroxyl radicals (*OH). These results were strong evidence of direct participation of *OH, formed by the Haber-Weiss reaction, in the water-soluble fraction of baked sponge cakes. The rate of decrease in active oxygen by scavengers decreased in Leghorn eggs more efficiently than in silky eggs. 4. The present experiments suggested that the use of silky fowl eggs could improve the quality and oxidative stability of baked cakes.

Antithrombogenic elastomers: novel anticoagulant/complement inhibitor-controlled release systems.

We have introduced novel synthetic anticoagulant and complement inhibitors for controlled release systems with high biocompatibility. These drugs were molecularly designed for extremely high biospecific inhibition, are readily soluble in polar organic solvents, such that they have versatile applications in commonly used hydrophilic polymeric systems via an easily attainable one-step co-casting technique at a given amount of loading. Another characteristic feature of the controlled release system is that both release rate and duration are controlled by the material, formulation and fabrication variables. These are easily manipulated by the hydrophilicity of polymers, amount of loading and film thickness. The drug-impregnated system may generate a new dimension in the formulation of controlled release biocompatibility.

Definition of the mitogenic factor (MF) as a novel streptococcal superantigen that is different from streptococcal pyrogenic exotoxins A, B, and C.

Human T cell activation by recombinant mitogenic factor (rMF) was investigated in comparison with that by recombinant streptococcal pyrogenic exotoxins (rSPE) A, B, and C. Recombinant MF, rSPEA, and rSPEC were mitogenic for peripheral blood mononuclear cells (PBMC), whereas rSPEB was not. Recombinant MF required only HLA-DR for the stimulation of PBMC, as determined using monoclonal antibodies (mAb) to HLA class II molecules and the mouse L cells transfected with HLA class II molecules. Recombinant SPEA and rSPEC required HLA-DR or HLA-DQ molecule. Recombinant MF selectively stimulated V beta 2, V beta 7, V beta 8, V beta 18 and V beta 21-bearing T cells, whereas rSPEA and rSPEC activated V beta 2 and V beta 6-bearing T cells as evaluated by the quantitative T cell receptor (TCR) analytical method. No clonality was observed in the nucleotide sequences of complementarity determining region 3 of TCR V beta in T cells responding to rMF. The profiles of cytokine production by PBMC in response to rMF, rSPEA, and rSPEC were quite similar. In summary, these results demonstrate that both HLA class II molecules and the TCR V beta required for rMF-mediated T cell activation are distinct from those required for rSPEA or rSPEC-mediated activation. Therefore, the MF is a novel streptococcal super-antigen which is different from SPEA, SPEB, and SPEC.

Establishment and characterization of an anti-idiotypic CD4+ CD8- T cell line to murine anti-alpha(1 --> 3) dextran antibody.

It is known that anti-alpha(1 --> 3) dextran antibodies of BALB/c mice are ordinarily of distinctive idiotypes (Id), one of which is the individual idiotype (IdI) that is represented by J558 or M104E to myeloma protein. In the present study, we established T cell line of Th1 type which recognized the Id of anti-alpha(1 --> 3) dextran antibody, and investigated its specificity and functions. The T cell line, named J-2R, had a phenotype of CD3+ CD4+ CD8- and expressed alphabeta-T cell receptors (TcR). J-2R proliferated in response to J558 in an I-Ed-restricted manner but did not respond to M104E which had substitution at amino acids 100 and 101. We confirmed that J-2R recognized J558 IdI, using synthetic peptides corresponding to two serial amino acid residues, Arg100 and Tyr101, spanning the J558 IdI in the third hypervariable region (hv3) of the heavy chain. alpha(1 --> 3) dextran-binding B cells which were isolated from dextran-immunized mice activated J-2R, but B cells from nonimmune mice did not. J-2R produced IL-2, IFN-gamma and IL-6, but did not produce IL-4, IL-5, or IL-10. Furthermore, J-2R inhibited the growth of J558 myeloma cells inoculated to the syngeneic mice in vivo. These findings suggest that Id-specific CD4+ T cells, J-2R, are involved in Id network and may play a role in vivo. J-2R is useful for analysis of the role of the Id-specific helper T cells in immune network because J558 IdI is frequently present on anti-alpha(1 --> 3) dextran antibodies.

Recognition of rheumatoid arthritis synovial antigen by CD4+,CD8- T cell clones established from rheumatoid arthritis joints.

OBJECTIVE: To investigate the rheumatoid arthritis (RA)-specific autoantigen(s) recognized by CD4+ T cells in patients with RA. METHODS: CD4+,CD45RO+ T cell clones were established from the joints of RA patients, and were examined for their proliferative response to synovial cells. RESULTS: Eight of 146 T cell clones responded to RA synovial cells in a DR-restricted manner. These T cell clones recognized solubilized antigens extracted from RA synovial cells in the presence of DR-matched antigen-presenting cells, but did not respond to those extracted from non-RA synovial cells. The antigens had a molecular weight of 50/25 kd. Five of the 8 T cell clones used T cell receptor BV6, and the remaining clones used BV12.2. CONCLUSION: The antigens recognized by joint-infiltrating CD4+ T cells are present exclusively in RA synovial cells. The expression of these antigens by synovial cells may trigger the autoreactivity of T cells in RA joints.

The AP-1 binding site in the feline immunodeficiency virus long terminal repeat is not required for virus replication in feline T lymphocytes.

Sequences of 31 bp containing putative AP-1 and AP-4 binding sequences in the U3 region of the feline immunodeficiency virus (FIV) long terminal repeat (LTR) were deleted and the basal promoter activity of the LTR was measured by the chloramphenicol acetyltransferase (CAT) assay. The activity of the FIV LTR was reduced in Felis catus whole foetus 4 (fcwf-4) cells and Crandell feline kidney cells by this deletion. Cotransfection of murine c-Fos or c-Jun expression plasmids with the FIV LTR-CAT reporter plasmid into fcwf-4 cells revealed that FIV LTR could be activated by c-Fos but not c-Jun in the cells. The mutated LTR was introduced into an infectious molecular clone of FIV and the replication rate and the cytopathogenic activity of the mutant were compared with those of the wild-type in two feline CD4-positive T lymphoblastoid cell lines. It was found that the rate and activity of the mutant were almost the same as those of the wild-type. From these data, we conclude that the 31 bp fragment is important for achieving maximal expression of the FIV genome, but not required for the replication of FIV in feline T lymphocytes.

Apoptotic cell death in arrhythmogenic right ventricular cardiomyopathy: a comparative study with idiopathic sustained ventricular tachycardia.

Arrhythmogenic right ventricular cardiomyopathy (ARVC) is a specific heart muscle disease of unknown etiology characterized by fatty and fibrofatty replacement of the right ventricular myocardium. It often manifests life-threatened ventricular arrhythmias. Previous studies have hypothesized that myocyte apoptosis contributes to the myocyte cell loss and fatty change in ARVC and may be induced by recurrent ventricular tachycardia (VT). We examined whether these progressive pathological changes result from apoptotic cell death in both autopsied and biopsied right ventricular myocardium from 35 patients with ARVC by using in situ terminal deoxynucleotidyl transferase assay (TUNEL) and agarose gel electrophoresis. We also studied the biopsied myocardium from 30 patients with idiopathic sustained VT whose origin was the outflow tract of the right ventricle. TUNEL-positive cells indicating DNA fragments were observed in some cardiomyocytes and fibroblasts in ARVC, but the numbers of TUNEL-positive myocytes were very low in idiopathic VT. DNA laddering was confirmed in two autopsied cases in ARVC, but not in a non-cardiac case who died. These results suggest that at least some cardiomyocytes and fibroblasts are subjected to apoptosis in ARVC, leading to the loss of myocardium with characteristic pathological changes and subsequently progressive cardiomyopathy. Furthermore, the apoptotic process may not result from myocardial ischemia due to repetitive VT.

Significant myocardial pathology and increase of alpha 1-adrenergic receptor number affecting the arrhythmogenic condition in cases with ventricular tachycardia.

To study the pathogenesis of idiopathic ventricular tachycardia with left bundle branch block morphology (LBBB-VT), histopathological findings in 34 patients and adrenergic receptor (AR) density obtained by micro-autoradiography in 11 patients were analyzed, using endomyocardial biopsy samples. According to the ventricular volume, we divided the patients into 3 groups. Group A patients (n = 8) were diagnosed as having arrhythmogenic right ventricular dysplasia (ARVD) showing right ventricular enlargement, group B patients (n = 7) showed decreased left ventricular function and/or enlarged left ventricle, and group C (n = 19) included patients with idiopathic ventricular tachycardia (VT) without enlargement or dysfunction of either ventricle. Histologically, there was a high incidence of significant pathology showing myocardial hypertrophy, degeneration, abnormal branching, and interstitial fibrosis in all groups (group A, 100%; group B, 86%; group C, 56%). There was a higher incidence of fatty tissue infiltration in groups A (100%), B (71%), C (48%) than in the control groups. As for AR, specific grains of alpha 1- and beta-AR were 23.0, and 18.3 (grains/25 x 25 mm square) respectively, in patients with LBBB-VT. The number of alpha 1 grains in patients with LBBB-VT was apparently higher than in the control group, sick sinus syndrome (SSS; 11.2), and the beta-AR density in the LBBB-VT group was the same as in the control group (SSS; 15.4). We concluded that the significant pathology in cases with VT might affect the arrhythmogenic condition. Moreover, these results suggested that the increase in the number of alpha 1-AR might have a great influence in idiopathic VT.

Establishment and characterization of nurse cell-like clones from human skin. Nurse cell-like clones can stimulate autologous mixed lymphocyte reaction.

We have established nurse cell-like clones from long-term cultures of the human skin. These human skin nurse cell (HSNC)-like clones were type I collagen+, type IV collagen-, vimentin+, cytokeratin-, CD44+, CD54+, and weakly positive for VCAM-1, and easily identified by the pseudoemperipolesis that allowed T lymphocytes to migrate beneath the HSNCs. HSNCs and various T cell lines formed a typical complex in the hanging drop culture system. The majority of human and murine T cells, and some of the tumor cell lines other than T cells, including B lymphoma and myeloblastoma cells, migrated beneath the HSNC clones. HSNC clones produced various cytokines, including IL-6, IL-7, IL-8, IL-9, granulocyte CSF (G-CSF), granulocyte-macrophage CSF (GM-CSF), macrophage CSF (CSF-1), TGF-beta 1, and c-kit ligand, but could not produce IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, TNF-alpha, or TNF-beta. These characteristics were similar to those of nurse cells established from the murine thymus. Furthermore, IFN-gamma-pretreated HSNC clones that expressed MHC class II Ags induced autologous mixed lymphocyte reaction (AMLR) in autologous PBMCs to proliferate and exhibit the cytotoxicity against altered autologous cells and various tumor cells. These results suggest that HSNCs play an important role in the immunoregulation at skin tissues.

[Radionuclide assessment of cardiac involvement in patients with sarcoidosis].

This study was aimed to determine whether nuclear methods were useful for examining cardiac pathology and for making a decision of corticosteroid therapy in patients with sarcoidosis. Thirty six patients were divided into two groups; GpA consisted of 19 patients with cardiac sarcoidosis and abnormal ECG findings, and GpB of 17 patients with sarcoidosis without ECG abnormalities. Cardiac uptake of 67Ga-citrate in 2 and 99mTc-PYP in one of GpA was observed and steroid therapy resulted in the disappearance of the uptake. 201Tl-CL cardiac tomograms disclosed perfusion defects in 10 of 14 patients (71%) in GpA, including defects with redistribution in 8 of the 10 pts, but only one case in GpB. Radionuclide ventriculography using 99mTc-RBC revealed abnormal response of left ventricular (LV) function to exercise and LV dysfunction in GpA. These data suggest that nuclear study is a useful tool for diagnosing cardiac sarcoidosis, evaluating therapeutic effectiveness and long-term follow-up in patients with sarcoidosis.

Feline CD4 molecules expressed on feline non-lymphoid cell lines are not enough for productive infection of highly lymphotropic feline immunodeficiency virus isolates.

To investigate whether the feline CD 4 (fCD4) molecules are involved in infections of highly lymphotropic feline immunodeficiency virus (FIV) isolates, we expressed fCD4 stably on Crandell feline kidney cells and Felis catus whole foetus 4 cells by transfection of a cDNA encoding the fCD4 glycoprotein, and then infected them with TM 1 and TM 2 strains of FIV, which are unable to infect these cells productively. In spite of fCD 4 being expressed on these cells, no virus production was observed. This result indicates that fCD 4 expression alone cannot induce a productive infection of the FIV TM 1 and TM 2 strains.