The effect of Cernitins on galactosamine-induced hepatic injury in rat.

Cernitins correspond to microbiologically fermented pollen extract. Cernitin T60 contains mainly water soluble, while Cernitin GBX mainly fat soluble substances. The aim of the present work was to examine the effect of Cernitins on the d-galactosamine-induced liver damage in rat. It has been shown, that galactosamine administration to rat resembles viral hepatitis both biochemically and histologically. Our studies proved, that Cernitin T60 given orally or intraperitoneally inhibited or counteracted the elevation of amino transferases activity and the inflammatory process, necrosis and steatosis of the liver cells. The protective effect of Cernitin GBX on the liver parenchyma was only slightly expressed. It is concluded, that application of pollen extracts in patients with liver diseases should be considered.

The protective effect of pollen extracts against allyl alcohol damage of the liver.

In male Wistar rats the hepatoprotective effect of pollen extracts (Cernitins) agains. orally introduced 1% allyl alcohol (0.4 ml per 100 g body weight) was investigated Cernitins were applied orally at 0.6, 24 and 30 h after allyl alcohol administration. After 48 h an autopsy was performed and blood was collected for biochemical tests. Liver damage was evaluated by measurement of aminotransferases (AspAT, AlAT) and alkaline phosphatase activity, total bilirubin level in the blood serum as well as by histological examination of the livers. Cernitins significantly reduced the serum enzymes elevations induced by allyl alcohol. The hepatoprotective properties of Cernitins were confirmed by histopathological studies.

The effect of short-term immunotherapy with Pollinex on anti-IgE and antigen-induced histamine release from sensitive human basophils.

Anti-IgE and antigen-induced histamine release from basophils isolated from 30 atopic patients sensitive to grass pollen allergens was evaluated. The studies were made before and after short-term immunotherapy with glutaraldehyde-pollen-tyrosine adsorbate (Pollinex). It was shown, that after immunotherapy a significant drop in anti-IgE and specific antigen-induced histamine release from isolated basophils occurs. The investigations confirm that the diminution of the target cell sensitivity and releasability may be one of the effects of specific immunotherapy.

Seasonal variation of in vitro lymphocyte proliferative response to ragweed antigen E.

Seasonal variation of symptoms and IgE response to short ragweed (SRW) allergens is well documented. Clinical symptoms generally parallel the rise and fall of the SRW-pollen count, whereas total and specific-IgE levels peak after the SRW-pollen season with a more gradual return to preseason levels. Because IgE synthesis is under T-lymphocyte control, we tested for seasonal variation in T cell-proliferative response to SRW antigen E (AgE) in vitro. Nine untreated SRW-sensitive and five nonallergic individuals were studied on 15 occasions from June 1981 through May 1982. In vitro proliferative index (SI) to AgE, serum total and specific IgE and SRW-pollen counts were measured; all persons studied kept daily symptom diaries. The mean SI was higher for the atopic group on all 15 sampling dates. The cumulative SI and the daily SI were statistically different between groups before, during, and after pollination. The peak SI for the atopic patients occurred almost 1 2/5 wk after the pollination peak, and the peak IgE antibody levels to SRW occurred at 5 2/5 wk after the pollination peak. We conclude that in vitro responsiveness to AgE is a specific response of allergic individuals and that this response demonstrates a significant seasonal component.

Further characterization and biologic activity of ragweed antigen--induced neutrophil chemotactic activity in man.

We have previously described the appearance in serum of increased neutrophil chemotactic activity (NCA) during bronchospasm induced by inhalation of ragweed antigen in ragweed-sensitive subjects. This NCA is non-complement derived, appears within 1 min after antigen inhalation, and is not seen after methacholine-induced bronchospasm. This article describes further characterization of this chemotactic activity and correlation with in vivo leukocytosis. NCA consistently eluted in the void volume (fraction I) after Sephadex G-150 chromatography of patient serum obtained 10 min postchallenge. Fraction I contained 94% of the NCA of postchallenge whole serum. Both postchallenge whole serum and fraction I deactivated neutrophils to autologous chemoattractants and complement-derived chemotactic factors, but not serum-independent chemotactic factors. NCA was chemotactic for neither human nor guinea pig eosinophils, nor for human mononuclear cells. A significant increase of circulating neutrophils was seen only after antigen-induced bronchospasm and correlated with the increase in NCA. Thus, NCA represents another inflammatory mediator of probable mast cell origin that may explain, at least partially, the accumulation of neutrophils observed in the peripheral blood, skin, and bronchial wall after immediate hypersensitivity reactions.

Influence of aeroallergens on bronchial reactivity in children sensitized to grass pollens.

Nine children, sensitized to grass pollens, with seasonal rhinitis and mild asthma underwent bronchial challenge with carbachol and EIA test repeatedly either during a pre-seasonal or in seasonal period. We failed to find any significant difference between pre-seasonal and seasonal values in both tests. We believe that atmospheric conditions are an important determinant of our results. It is our hypothesis that a threshold concentration in atmospheric pollen may exist which must be reached before an allergic reaction is evident.

Changes in the electroencephalographic and gamma-aminobutyric acid transaminase and succinic semialdehyde dehydrogenase in the allergen induced rat brain.

Electroencephalographic activity and gamma-Aminobutyric acid Transaminase together with Succinic semialdehyde dehydrogenase activity changes produced by sensitization with Prosopis juliflora pollen allergen were studied in the cerebral cortex and hypothalamus of the rat brain. Electrical activity of EEG recording begins to appear on 3rd day after sensitization with maximum increase in activity was found on day 9 and decreased after that. A sudden increase in electrical activity was produced in 9th day sensitized rat with 10 min after giving challenging dose intravenously. The measurement of enzymatic activity of GABA-T and SSA-DH showed decrease and increase in 3, 9, 15 and 30 days sensitized rat hypothalamus and cerebral cortex whole homogenate and mitochondrial fractions. A maximum changes in enzymatic activity was found in 9th day sensitized rat with significant alterations after giving sudden stress as challenging dose. These changes in EEG activity and GABA-ergic neurotransmitter in allergenic rats showed the immunoregulatory role of nervous system mediated via GABA shunt.

Induction of allergen-specific T cells by conjugates of N-formyl-methionyl-leucyl-phenylalanine and rye grass pollen extract.

A conjugate of the biologically active peptide N-formyl-methionyl-leucyl-phenylalanine and rye grass pollen extract (F-MLP/rye), previously shown to react with rye grass pollen extract-specific T cells, induced the formation of allergen-specific T cells in mice. Lymph node cells prepared from mice immunized with either native extract or F-MLP/rye gave an enhanced response to unmodified rye grass pollen allergens in vitro. Syngeneic spleen macrophages were able to present the unmodified allergens to T cells obtained from both groups of mice causing their proliferation in vitro. Conjugation of the peptide into the extract brought about an extensive reduction in its reactivity with grass pollen-specific human IgE, and a loss of its ability to induce specific IgG antibody in guinea-pigs. A state of delayed hypersensitivity specific for rye grass pollen extract was produced in guinea-pigs by immunization with either the F-MLP/rye or unmodified extract. It is concluded that conjugates such as F-MLP/rye or other T' allergoids could be used as probes to investigate whether changes in T cell activity are important in immunotherapy.

[Inhibitory effect and synergism of cernitin pollen extract on the urethral smooth muscle and diaphragm of the rat].

Inhibitory effects of cernitin pollen extract (CN-009), consisting of T-60 (a water-soluble extract) and GBX (an acetone-soluble extract) at a ratio of 20:1, were investigated in rat urethral smooth muscle and diaphragm. In the urethral smooth muscle, CN-009 (3.0 x 10(-4) approximately 3.0 x 10(-3) g/ml), T-60 and GBX (corresponding to CN-009) concentration-dependently inhibited the noradrenaline (NA, 10(-5) g/ml)-induced contraction. According to Bürgi's calculation, the inhibition by T-60 and GBX was synergistic. On the other hand, GBX inhibited the NA-contraction non-competitively and also inhibited the K+-contraction. In contrast, T-60 tended to inhibit competitively, but did not affect the K+-contraction. In the diaphragm, CN-009 (5.25 x 10(-3) approximately 2.1 x 10(-2) g/ml) concentration-dependently inhibited the nerve stimulation-induced twitch response. T-60 (corresponding to CN-009) showed no effect, while GBX slightly inhibited the twitch response. The effects of T-60 and GBX were synergistic. The inhibitory effects of CN-009 (2.1 x 10(-2) g/ml) and GBX (1.0 x 10(-2) g/ml) were specific against the nerve stimulation and were not antagonized by neostigmine (1.0 x 10(-5) g/ml). These results suggested that these effects were neither musculotropic nor competitive against ACh. In conclusion, CN-009 concentration-dependently inhibited the urethral contraction and the skeletal muscular twitch response. It was suggested that T-60 and GBX had different mechanisms and inhibited the response synergistically.

Lymphocyte proliferation to antigen E: demonstration of the restriction of antigen E-specific T cells to ragweed-allergic donors.

Allergic sensitivity to ragweed is common among atopic individuals in North America and can be associated with symptoms of seasonal hay fever and increased airway reactivity in asthma. This sensitivity is mediated by IgE antibody to ragweed antigens that in turn is presumed to be the product of B-lymphocytes regulated by various T cell subsets. Proliferation in vitro by lymphocytes obtained from individuals allergic to ragweed and cultured in the presence of ragweed antigen E (AgE) has been repeatedly described, but a comprehensive study of this proliferation has questioned the specificity of this response. We have examined this question and found that in the first week of culture, the specific lymphocyte proliferation to AgE may be obscured by high background and mitogen-like proliferation. However, by carrying the cells for a longer period of time in culture and providing a second in vitro boost with AgE, specific proliferation could be clearly documented. Lymphocytes from atopic ragweed-allergic donors proliferated at levels 20 to 50 times beyond background in the presence of AgE. Cells from nonragweed-allergic donors (either nonatopic or atopic) did not do so. The AgE-responsive cells could be expanded in culture and demonstrated to be T cells. Moreover, AgE-responsive T cells could only be cloned from AgE-allergic donors and, after expansion and subcloning, demonstrated to respond to AgE but not partially purified dust mite antigen. In contrast, a clone of T cells from a dust mite-sensitive individual proliferated in response to the dust mite antigen but not AgE.

Modulation of histamine release from human basophils in vitro by physiological concentrations of zinc.

Zinc, at physiologic concentrations, inhibits in vitro histamine release from human basophils induced by several immunologic (i.e., antigen and anti-immunoglobulin E (IgE) and nonimmunologic [Ca++ ionophore A23187 and formylated tripeptide formyl-methionyl-leucyl-phenylalanine (f-met peptide)] stimuli in a dose-dependent manner. Inhibition begins at about 10(-6) (ionophore A23187, anti-IgE and antigen) or 10(-5) M (f-met peptide) and is maximum at 10(-4) M (80--100% inhibition of histamine release). The activity of zinc is about 25-fold greater with respect to ionophore A23187 (ID50 = 1.1 x 10(-6) M) than to f-met peptide-induced (ID50 = 4 x 10(-5) M) histamine release. Its activity on IgE-mediated histamine release is intermediate between these two extremes (ID50 = 9.7 x 10(-6) M). Zinc does not affect the first stage of histamine release but acts on the calcium-dependent second stage. It is a competitive antagonist of the action of Ca++ in histamine secretion induced by antigen, anti-IgE and f-met peptide (but not by A23187) with a dissociation constant of about 1.2 x 10(-5) M. The addition of colchicine with zinc fails to increase the inhibition caused by the ion alone, suggesting the two compounds work via a common mechanism of action. Deuterium oxide reversed, in a dose-dependent manner, the inhibition of histamine release caused by zinc. These results suggest that the effect of zinc on histamine release from human basophils may be related to its influence on the microtubule system, directly or via its interaction with calcium.

Metabolic adaptation of muscles to exercise, vibration and raised temperature under the influence of cernitins.

Wistar rats were used to study the effects of cernitins, i.e. aqueous and oil extracts of pollens, on the metabolic adaptation of the soleus muscle to exercise, vibration and raised ambient temperature. The animals were exposed to selected combinations of these factors for 5 days during 1.5 hour daily. A part of the animals was given orally cernitins in daily doses of 6 mg/kg of body weight for 10 days before the exposure. Among the adaptation changes studied in the soleus muscle, 24 hours after the last exposure, cernitins caused: a reduction of the amount of total protein, an increase in the proportion of soluble proteins in the protein fraction, an increase in the tissue oxygen consumption, an increase of already elevated pyruvate kinase activity, a further rise in ATP level, an increase in lactic dehydrogenase activity, a rise in the activity of cholinesterases. Moreover, it increased significantly the body weight and the weight of the studied soleus muscle. Cernitins, in combination with certain types of exposure used in this experiment, exerted a catabolic action, increased the rate of anaerobic metabolism and enhanced adaptation to exercise, vibration and temperature. The direction of the adaptation changes depended on the type of exposure to environmental factors.