Rabbit autoantibodies to actin induced by immunization with heterologous actins; a possible mechanism of smooth muscle antibody production

Antisera against actins from chicken gizzard smooth muscle and ascaris body wall were prepared in rabbits. Immunological cross-reactivity of the antisera with actins of several different species was demonstrated by precipitation reactions in agarose gels and by immunofluorescence studies. The antisera were also reactive with actin of rabbits, the homologous animal used for immunization. The latter finding indicated termination of natural tolerance to actin by immunization of cross-reactive actins. A possible mechanism operating in the induction of smooth muscle antibodies, autoantibodies which have a similar nature to the anti-actin antibody is discussed in relation to the termination of natural tolerance to actin.     

Mechanoenergetics characterizing oxygen wasting effect of caffeine in canine left ventricle

Caffeine causes a considerable O(2) waste for positive inotropism in myocardium by complex pharmacological mechanisms. However, no quantitative study has yet characterized the mechanoenergetics of caffeine, particularly its O(2) cost of contractility in the E(max)-PVA-VO(2) framework. Here, E(max) is an index of ventricular contractility, PVA is a measure of total mechanical energy generated by ventricular contraction, and VO(2) is O(2) consumption of ventricular contraction. The E(max)-PVA-VO(2) framework proved to be powerful in cardiac mechanoenergetics. We therefore studied the effects of intracoronary caffeine at concentrations lower than 1 mmol/l on left ventricular (LV) E(max) and VO(2) for excitation-contraction (E-C) coupling in the excised cross-circulated canine heart. We enhanced LV E(max) by intracoronary infusion of caffeine after beta-blockade with propranolol and compared this effect with that of calcium. We obtained the relation between LV VO(2) and PVA with E(max) as a parameter. We then calculated the VO(2) for the E-C coupling by subtracting VO(2) under KCl arrest from the PVA-independent (or zero-PVA) VO(2) and the O(2) cost of E(max) as the slope of the E-C coupling VO(2)-E(max) relation. We found that this cost was 40% greater on average for caffeine than for calcium. This result, for the first time, characterized integratively cardiac mechanoenergetics of the O(2) wasting effect of the complex inotropic mechanisms of intracoronary caffeine at concentrations lower than 1 mmol/l in a beating whole heart.     

Non-IgG1 nature of cutaneous basophil hypersensitivity factor in contact sensitivity. II. Demonstration of antigen-dependent basophil chemotactic activity of cutaneous basophil hypersensitivity factor

It was recently demonstrated that there was a specific activity to induce basophil-rich skin reaction in the sera of contact-sensitized guinea pigs (CBH factor, CBH-F). In the in vitro chemotactic assay system, CBH-F was shown to have weak basophil chemotactic activity but enhanced its activity in the presence of corresponding antigen(s). Furthermore, basophil chemotaxis in reaction to the antigen(s) was observed when the cells were preincubated with CBH-F. It worked mainly for Percoll-separated bone marrow basophils but not for oil-induced peritoneal macrophages. Immunological analysis revealed that CBH-F was a protein with a small molecular weight (MW 4-6 X 10(4] with an antigen binding site and isoelectric point of between 4.5 and 5.0. It did not show any characteristics of IgG1 or IgG2 on immunoadsorbent column and immunoelectrophoresis. Enzyme treatment with insoluble trypsin eliminated the chemotactic activity but this was not the case with neuraminidase treatment.     

Contact sensitivity in vitro: The production of macrophage inhibition factors from DNCB sensitized lymphocytes by subcellular organelles obtained from DNCB epidermal tissue

Actively allergized cells from guinea-pigs with contact sensitivity to DNCB were specifically triggered to produce the macrophage inhibition factors—antigen-dependent and non-antigen-dependent MIF by microsomal—DNCB complexes. The implication of this observation is that epidermal microsomal—DNCB complexes behave as complete antigens in a MIF assay system. It is therefore possible that such a combination acts as an immunogen in the induction of contact sensitivity to DNCB.     

Novel centrifugal method for simple and highly efficient adenovirus-mediated green fluorescence protein gene transduction into human monocyte-derived dendritic cells

Dendritic cells (DC) are professional antigen-presenting cells in the immune system. Gene transduction of DC with tumor-associated antigen (TAA) or other genes that enhance the immune reaction has been considered theoretically useful for DC-based immunotherapy. However, gene transduction of DC generated from human peripheral blood monocytes has been difficult due to its low efficiency, even when adenoviral vector was used at high multiplicity of infection (MOI). In the present study, we examined the effect of centrifugal force to enhance efficiency of adenovirus-mediated gene transduction into human monocyte-derived DC at various rotor speeds at various temperatures for various times. We judged the transduction efficiency using enhanced green fluorescence protein (EGFP)-expressing adenoviral vector, and the best condition for centrifugal transduction was determined as 2000 x g at 37 degrees C for 2 h at an MOI of 10 or greater. At an MOI of 50 without centrifugation, the gene transduction efficiency was about 66% and mean fluorescence intensity (MFI) of EGFP expression was about 150 (at 37 degrees C for 2 h). With centrifugal transduction (2000 x g at an MOI of 50 at 37 degrees C for 2 h), 86% or more DC were gene-modified, and especially, MFI of EGFP expression was highly enhanced (MFI: about 3100 or greater). Centrifugally gene-transduced DC were not damaged and were thoroughly functional as measured by mixed lymphocyte reaction (MLR). The centrifugal method was also applicable to human monocytes and K562 cells. The centrifugal transduction method with adenoviral vector might be helpful for the generation of gene-modified DC.     

Molecular cloning of cDNAs expressing SS-B/La protein

Using serum from a patient with Sj?gren's syndrome containing a high titer of anti-SS-B/La antibody, cDNA clones (a representative clone was called pA158) were isolated from a human fibroblast cDNA library in lambda gt11 expression vector. After subcloning of pA158 cDNA into an expression plasmid vector pEX-2, a large amount of the recombinant fusion protein with cro-beta-galactosidase (called pA158EX) was obtained in E. coli culture containing the recombinant pEX-2. Antibodies against pA158EX were purified from the patient serum by Sepharose 4B conjugated with the purified pA158EX protein. Immunofluorescent staining of HEp-2 cells with the anti-pA158EX antibodies showed a speckled nuclear staining. In immunoblot analysis, the anti-pA158EX antibodies reacted with 50 kDa protein that was compatible with SS-B/La protein. Immunoprecipitation of leukocyte lysate with the anti-pA158EX antibodies and the following RNA analysis showed that the antibody precipitated Y5 RNA. These findings indicate pA158 is a cDNA for SS-B/La protein. The purified fusion protein was used for enzyme-linked immunosorbent assay (ELISA). Optical density values of anti-SS-B positive sera were high, but those of anti-SS-B negative sera and healthy donor sera were low. In the Northern blot using human RNA and pA158 cDNA, a single band about 1.8 kb was recognized. A full-length cDNA was further obtained by screening of pcD library using pA158 cDNA as a probe.     

Clinical experience with ceftizoxime suppositories in pediatrics

Ceftizoxime suppositories (CZX-S), containing 250 mg or 125 mg of CZX, were given to 6 children, 4 with acute bronchopneumonia and 2 with acute pharyngobronchitis, who were not suited to treatment with injectable or oral form of the drug. The clinical response was "good" in all the children and the causative organisms were eradicated in 2 children (H. influenzae or S. aureus). Adverse reactions consisted of 1 case each of diarrhea and transiently increased GPT. In conclusion, CZX-S proved to be highly effective in the treatment of bacterial infections in children.     

Safety and efficacy of Tolvaptan in real-world patients with autosomal dominant polycystic kidney disease- interim results of SLOW-PKD surveillance

Clinical and Experimental Nephrology - Tolvaptan is a vasopressin type 2 receptor antagonist and has been used to treat autosomal dominant polycystic kidney disease (ADPKD) since 2014. There has...