Iron chelation effects on lipid peroxidation, inflammation and ventricular performance in a rat model of isoproterenol induced acute myocardial stress.

Acute cardiac pathologies represent one of the leading causes of death, while iron metabolism is recognized to be implicated in reactive oxygen species production, lipid peroxidation, and inflammation. The aim of the present study was to assess iron chelation effects in isoproterenol (ISO) induced acute cardiac stress. We divided male Wistar rats into preventive and secondary treatment groups, with the active arm consisting in deferiprone (DFP), a lipid permeable chelator. Mortality of ISO was 10-18.18% in both preventive and secondary groups. We analyzed serum and myocardial tissue parameters of inflammation, iron dynamics, and lipid peroxidation, accompanied by ultramicroscopy, histological, and ultrasound-derived parameters of left ventricular function. Results reveal that ISO-mediated lipid peroxidation and inflammation are alleviated by administration of DFP, with negligible effect on systemic ferroregulation dynamics and global ventricular function (as assessed by ultrasound). DFP administration after cardiovascular stress is associated with a decrease in lipid peroxidation and inflammation, without an improvement in gross left ventricular parameters.

Characterization and biological effects of Hypericum extracts on experimentally-induced - anxiety, oxidative stress and inflammation in rats.

Anxiety disorders can associate with oxidative stress and immune system alterations. Our study aimed to chemically analyze Hypericum maculatum (HM) and Hypericum perforatum (HP) dry extracts and to evaluate their effects along with quercetin (Q), on brain oxidative stress biomarkers: malondialdehyde (MDA), catalase (CAT) and superoxide dismutase (SOD), inflammatory cytokines: interleukin-1α, (IL-1α), IL-1ß, regulated on activation normal T cell expressed and secreted (RANTES), interferon (IFN), monocyte chemoattractant protein-1 (MCP1), macrophage inflammatory protein (MIP) and serum corticosterone levels. Nuclear transcription factor κB (NFκB) signaling pathway in the hippocampus and frontal lobe in rats with N-methyl-9H-pyrido[5,4-b]indole-3-carboxamide (FG-7142) experimental-induced anxiety were also investigated. The chemical analyses of total hypericins were performed by spectrophotometric analysis and hypericin, hyperforin and polyphenols derivatives were quantified by chromatographic methods. The animals were divided in 6 groups: carboxymethylcellulose 2% (CMC); CMC + FG; alprazolam (APZ) + FG; Q + FG; HM + FG; HP + FG. APZ (0.08 mg/kg b.w.), Q (30 mg/kg b.w.), HM and HP (350 mg/kg b.w.) were orally administered for 21 days. FG (7.5 mg/kg b.w.) was intraperitoneally (i.p.) injected in a single dose. Q and hypericum extracts (HpE) exerted anti-inflammatory (decreased IL-1α, IL-1ß, MCP1, IFN and MIP mainly in hippocampus) and antioxidant effects (decreased MDA levels, increased CAT and SOD activity), enhanced NFκB and pNFκB expressions in the brain and reduced serum corticosterone levels. Our findings suggest that HpE may improve anxiety-like behavior, offer brain protection by modulation of oxidative stress and inflammation, and can contribute to overall biological activity of natural compounds-rich diet.

I. Immunosurveillance and tumorigenesis in 1. BALB/c mice. Early reactive state in the jejunum disseminated lymphoid tissue.

Using functional ultrastructure we established the physiology of the cellular immune response in the BALB/c mouse jejunum disseminated lymphoid tissue (JDLT) before and after sequential stimulations with E. coli endotoxin (ECE) (the preceding article in this journal). The functional ultrastructure was established also in this series of BALB/c mice injected i.p. initially with 0.5 ml of mineral oil and then weekly with 5 ng of ECE (EOA). The present series of mice had a pathological immune response and, it is known that they will in time produce plasmacytomas. Differences between the physiologic and pathologic immune responses during the first month of tumorigenesis, are: 1. The T-lymphocyte response was found to be strong but disorganized; 2. As judged from the T:B ratios, the B-lymphocyte response was found to be very strong, late after the first, early after the second, and late after the fourth injections with ECE, in this EOA model of stimulation. 3. The macrophage cellular response during the EOA stimulation was found to be slightly weaker than that seen in the series of mice stimulated with ECE alone. At the time of this study, the differences found between the physiologic and pathologic immune responses in the JDLT are not conclusive and they do not elucidate either the mechanisms of tumorigenesis or the role of the immunocompetent cells.

The in situ cellular immune response in BALB/c mice jejunum disseminated lymphoid tissue after stimulations with E. coli endotoxin.

The feasibility of the functional ultrastructure, a term used to describe the in situ immunocumpetent cell topography, B-cell transformation, multiplication and cell-to-cell interactions, was only recently made possible (Toma et al. 1977a; Toma and Retief 1978). The functional ultrastructure of the BALB/c mice jejunum disseminated lymphoid tissue (JDLT) before and after four i.p. sequential stimulations with 5 ng E. coli endoxin (ECE) each revealed the following results: 1. The in vitro T-cell response was found to be by recruitment and not by the in situ cell division. T-lymphocyte recirculation in the thymus may be the answer to the question "Where does the clonal proliferation occur?" 2. Two T-cellular immune responses have been identified, one occurring early during the stimulation (1st and 2nd stimulations) and the other occurring late (after the 4th stimulation with ECE). It was thought that the first T-cell response represented the helper type of the immune response, whereas the second would represent the suppressor T-cell response. 3. The B-cell response, as judged from the percentage of plasma cells (PC) generated, was strong in the T-PC islands but no patterns of its generation could be established. 4. The curious topography of the immunocompetent cells inside the T-PC islands in the Lieberkühn crypts was found and their origin discussed. We aimed to establish the physiology of the cellular immune response in JDLT and to use it as a normal comparative model. It will be compared with the pathology of immune response in this part of BALB/c mouse gut established by immunosurveillance and tumorigenesis in artificially induced plasmacytomas with mineral oil (the following article).

Plasma cell leukaemia. Diagnostic problems in our experience with 11 cases.

11 patients with plasma cell leukaemia (PCL) are reported. Diagnostic clinical, haematological, immunological, biochemical and electron microscopical (TEM) data were analysed and compared to the largest series of PCL cases reported in the literature. Special attention was paid to four facets of this disease: (a) the clinical picture at admission; (b) the frequency of PCL; (c) the production of M components in relation to the maturity and type of the asynchronous plasma cells, and (d) the diagnostic problems of this entity of acute leukaemia of the afferent limb of the B lymphocyte transformation. In this series PCL emerges as a distinct clinical entity: patients are severely anaemic, hepatosplenomegaly is prominent, bone lessions are uncommun, but if present are usually non-osteolytic, and the response to treatment with an alkylating agent and glucocorticoid is poor. The diagnosis is difficult since the circulating plasma cells may have morphological features which only allows the diagnosis to be made after the TEM examination. If the peripheral blood of cases of acute leukaemias and immunocytic dyscrasias is routinely examined by TEM, PCL appears to be a not uncommon variant of plasma cell dyscrasia--in the present study it was 11%.

Thrombocytopenic purpura in pregnancy. A case report.

A case of immune thrombocytopenic purpura in pregnancy is reported. An interesting facet of the treatment was the suppression of spontaneous labour while platelet concentrate was being prepared. This case clearly demonstrated that the maternal antibody titre in no way reflects the fetal status. The choice of delivery can therefor not be based upon scientific data. The possible use of the uranyl-labelled antibody method of determining antiplatelet antibodies may in future help to determine the best method of delivering these babies. A review of the literature clearly indicates that while maternal mortality has decreased, fetal loss in this condition is still significant, and that a multicentre study is needed to examine the best approach to this problem.

Is the d an allelic gene of D in the Rh blood group system? A quantitative study using 99mTechnetium-pyrophosphate-labelled antibody (TcLA) method.

This article reports on studies of Rh antigens, such as D, Du and d, using uranyl-labelled antibody (ULA) and TcLA (99mTc-pyrophosphate-labelled antibody) methods for the first time for this purpose. TcLA method proved to be simple in labelling and very sensitive (20--100 times more so than the indirect Coombs test) in the detection of Rh antigen-antibody binding. Results of this quantitative study demonstrate convincingly that the d is not an allelic gene of D but rather the weakest of the series D less than Du less than d. Although the evidence from this study demonstrates clearly that differences between D and d are only quantitative, the authors do not think that the Rh nomenclature should be changed but they do think that the present evidence should be used in regard to the understanding of the allelism in the Rh blood group system. The c is an allelic gene of C as the e is an allelic gene of E; specific test sera detecting every one of these antigens exist and the family studies verify these statements. However, the d is not a distinct antigen as c and e are, even if the pattern of inheritance from family studies, using the existent anti-D serum, would suggest the allelism as probability. That is why in the past the anti-Du and anti-d specific test sera never incidentally found or artificially produced.

Uranyl-labelled antibody (ULA) method and its application for Rh antibodies.

The existent labelling materials for studies of antigen--antibody interaction at ultrastructural level, namely ferritin and peroxidase, because of their large molecular size do not fulfill all requirements of excellent markers for electron microscopy (EM). Uranyl acetate has a molecule 354 times smaller than IgG and its uranium atom is electron-dense. These physical characteristics of uranyl acetate make it a labelling material par excellence as described in this article. Quantitative and qualitative studies of Rh antigen-antibody interactions are for the first time presented at the ultrastructural level, and the application of the uranyl-labelled antibody (ULA) method for weak antisera (dilutions 100 to 1000 time higher than the Coombs range of sensitivity) is demonstrated. The ULA method opens a new era for studies of antigens, antibodies and their interactions because it will demonstrate visibly details of the antigen-antibody interaction and is especially suitable for studies of weak antisera.

The identification of human T and B lymphocytes by electron microscopy.

Lymphocytes of peripheral blood, bone marrow, spleen, vermiform appendix, tonsil and Mantoux skin reaction were examined by electron microscopy (EM) and classified as T, B and Null cells by E-rosette and immunoglobulin membrane-receptor characteristics. The low pH and ionic strength of the fixative solution for EM, and some other minor procedural modifications, made it possible to distinguish B and T lymphocytes morphologically. T-cells have electron-dense cytoplasm and euchromatin in the nucleus whereas B-cells constantly have electron-lucent cytoplasm and euchromatin in the nucleus. A proportion of lymphocytes were unclassifiable by their ultrastructural features. These unclassifiable cells may be Null cells as determined by the classical techniques. The specificity and simplicity of this EM technique for T and B lymphocytes is especially useful for studies of immunocompetent-cell topography and cell-to-cell interaction in lymphoid organs. It may also be utilized for diagnostic purposes in immunocytic dyscrasias.