In vitro immunomodulatory activity of verbascoside from Nepeta ucrainica L

Nepeta ucrainica L. is used as a herbal tea in Kazakhistan. Phytochemical investigations of the aerial parts of the plant resulted in the isolation of verbascoside (1) and 1,5,9-epi-deoxyloganic acid (2). The structures of the compounds were elucidated by spectral (UV, IR, (1)H-NMR and (13)C-NMR) methods and HPLC analysis. The in vitro immunomodulatory activity of verbascoside was investigated by assessing neutrophil function; chemotaxis and intracellular killing activity. Verbascoside showed an increased chemotactic activity in all doses applied compared with the medium used as a negative control and had a positive effect on respiratory burst of neutrophils, but there was an opposite effect with increasing doses, pointing to a possible suppression of neutrophil intracellular killing activity.     

A study on geno- and immunotoxicological effects of subacute propoxur and pirimicarb exposure in rats

A 28-day oral exposure with 8.51, 3.40, and 0.851 mg/kg propoxur (PR) and 4.67, 1.87, and 0.467 mg/kg pirimicarb (PI) was performed in male Wistar rats, and the occurrence of numerical and structural chromosome aberrations and the changes in certain immune function parameters (plaque-forming cell (PFC) assay, delayed-type hypersensitivity reaction) and in some basic toxicological (body weight gain and weights of brain, thymus, lung, heart, liver, spleen, kidneys, adrenals, and popliteal lymph node) and hematological (white blood cells, red blood cells, hematocrit (Ht), mean cell volume of red blood cells (MCV) cell content of the femoral bone marrow) parameters were investigated. The high dose of PR increased the relative liver weight and the cell content of femoral bone marrow, and all three doses increased Ht and MCV. The applied doses of PI decreased the relative adrenal weight in a dose-dependent manner, and its highest dose increased the relative liver weight. Among the immune function parameters tested, PFC content of the spleen was decreased by high-dose PR and elevated by high-dose PI, whereas the maximum and the time course of the delayed-type hypersensitivity reaction showed no changes in this dose range. In the genotoxicological investigations only the high PR dose increased the number of numerical, but not the structural, chromosome aberrations. In addition to the changes in relative adrenal weight following PI treatment, the PFC assay showed the highest sensitivity for detection of the 4-week exposure with these carbamates. On the basis of our results, the immunotoxicological approach seems to have the same (PR) or higher (PI) sensitivity in early detection of the repeated low-dose exposure by these carbamates compared to the genotoxicological approach.     

Immunotoxicological effects of repeated combined exposure by cypermethrin and the heavy metals lead and cadmium in rats

The immunotoxic effect of a 28 days oral exposure by 55.4, 22.2, and 11.1 mg/kg cypermethrin (CY) was investigated in 4 weeks old male Wistar rats. The applied test system involved the determination of general toxicological parameters (body weight gain, organ weight of thymus, heart, lung, liver, spleen, kidneys, adrenals and the popliteal lymph node), haematological parameters (white blood cell count, red blood cell count, haematocrit, mean cell volume of red blood cells, cellularity of the femoral bone marrow), as well as immune function assays (splenic plaque forming cell assay, delayed type hypersensitivity reaction). The highest dose resulted in a significant increase of the relative liver weight, and all three doses resulted in (although inconsistent) changes in the haematocrit and MCV values. The maximum of DTH reaction decreased at all three doses. On combination of the highest CY dose with non-effective doses of lead or cadmium the immunotoxic effects of the former were modified. When immunotoxic doses of Cd or Pb were combined with the lowest CY dose, further interactions were observed on the examined parameters. The alterations of the immunotoxic effects of CY by simultaneous exposure with Cd or Pb, as described here, can lead to unexpected health consequences and/or can lead to false positive or negative results in human epidemiological studies.     

Simultaneous geno- and immunotoxicological investigations for early detection of organophosphate toxicity in rats

Detectability of toxic effects by repeated doses of dimethoate (DM) and methylparathion (MPT) were investigated by geno-and immunotoxicological methods in male Wistar rats following a 28-day oral exposure. In the dose range of 28.2, 14.1, and 7.04, and 7.04 mg/kg/day DM, the two higher doses decreased the body weight gain. The top dose increased the weight of liver, kidneys, and testicles; the white blood cell count; and the cell content of the femoral bone marrow. From immune function parameters measured [IgM-plaque forming cells (PFC) assay, delayed-type hypersensitivity (DTH) reaction] only the maximum of the DTH reaction decreased at the top dose. Of the MPT doses (0.872, 0.436, and 0.218 mg/kg/day) the two higher ones increased the liver weight, and a dose-dependent increase was found in the MCV value. No evaluable changes in the examined immune function parameters were observed. Both substances increased the number of numerical but not the structural chromosome aberrations at lower dose levels (the two larger doses of DM, and all the three doses of MPT) than those ones which caused changes in the examined immune function parameters. According to these results, the genotoxicological approach seems to be more sensitive for detection of repeated-dose oral toxicity of the investigated two organophosphates than the immunotoxicological one.     

Immunotoxicological examination of repeated dose combined exposure by dimethoate and two heavy metals in rats

The immunotoxicity of 28 days combined oral exposure by dimethoate (DM) and two heavy metals (Pb or Cd) was investigated in male Wistar rats. Immunotoxic and no-effect doses of DM (28.2 and 7.04 mg/kg) were combined with immunotoxic and no-effect doses of CdCl2 (6.43 and 1.61 mg/kg) or lead acetate (80.0 and 20.0 mg/kg) in such a way that the high dose of each substance was given in combination with the no-effect dose of the other. To examine the interactions of these agents, general toxicological (body weight gain, organ weights), haematological (absolute and differential WBC, RBC, MCV, Ht. cell content of the femoral bone marrow), and immune function (splenic PFC number. DTH reaction) parameters were measured. Treatment with the combination of Pb or Cd and DM did not result in a reduction of humoral (PFC) and cellular (DTH) immune responses, whereas treatment with the substances alone did result in immune suppression. This protecting effect can probably be attributed to an effect on the kinetics of the compounds tested rather than on the immune system itself. Further interactions were found in both combinations, DM-Cd and DM-Pb, in the body weight gain and in the relative liver weight; the DM-Pb combination also affected the relative thymus weight and the MCV value. These findings show that the immunotoxic effects of the investigated materials, including their detectability and health consequences, can be modified in case of combined exposure.     

Immunotoxicological investigation in rats dosed repeatedly with combinations of cypermethrin, As(III), and Hg(II)

The effects of the combined exposure of cypermethrin (CY), NaAsO2 (As), and HgCl2 (Hg) were investigated in male Wistar rats following 4 weeks of oral exposure. Standard toxicological (body weight gain, organ weights of brain, thymus, heart, lung, liver, kidneys, adrenals, testes, and popliteal lymph node), haematological (white blood cell, red blood cell, haematocrit, MCV-mean volume of red blood cells, and cell content of the femoral bone marrow), and immune function (IgM-PFC response, delayed type hypersensitivity (DTH) reaction) parameters were measured. The two doses selected for the combination were: a high (H) dose equal to lowest observed effect level determined in preliminary experiments (CY(H)=55.4 mg/kg, As(H)=13.3 mg/kg, Hg(H)=3.20 mg/kg), and a low (L) dose which was ineffective (non-observed effect level); CY(L)=11.1 mg/kg, As(L)=3.33 mg/kg, Hg(L)=0.40 mg/kg). CY(H) was combined with As(L) or Hg(L), and the H doses of the heavy metals were combined with CY(L). Beside vehicle control, the H dose components of the combinations were also used as internal controls. The main finding of this study was that certain combinations, when compared to H dose internal control, significantly altered the body weight gain (As(H)+CY(L)), the relative weight of adrenals and popliteal lymph node (CY(H)+As(L) and CY(H)+Hg(L)), the cell content of the femoral bone marrow (CY(H)+Hg(L)), and the time course of DTH reaction (CY(H)+As(L)). According to the present results, combined exposure with CY and the heavy metals investigated can modify the toxicity and/or the functional detection limit of the single substances.     

Preferential immunoglobulin oxidation in children with juvenile idiopathic arthritis

OBJECTIVE: Juvenile idiopathic arthritis (JIA) is a rare chronic inflammatory disorder of the joints. There is strong evidence that oxidative damage occurs in rheumatoid diseases, including JIA. The increased level of protein oxidation products in total plasma proteins has recently been reported in children with diagnosed JIA. The objective of this study was to find out which fraction of plasma proteins is mostly damaged by oxidative stress and whether the damaging effect correlates with certain clinical or laboratory parameters. METHODS: A new approach to estimate the carbonyl content of plasma protein fractions was developed, based on two-stage electrophoresis and immunochemical detection of the carbonyl derivatives of the proteins. This method allowed us to detect and quantitate carbonyl groups in the albumin, alpha-2, beta and gamma-globulin fractions. Sera of 25 children with JIA and 13 healthy controls were tested. RESULTS: Albumin and gamma-globulins were found to be most modified by oxidation. In a group of children with systemic JIA, both albumin and gamma-globulins were oxidized while plasma gamma-globulin fraction damage was prevalent in pauciarticular JIA. CONCLUSIONS: Among plasma proteins of children with JIA, gamma-globulins were preferentially oxidized, whereas most of the other proteins did not seem to be affected. Oxidative modification of plasma proteins was correlated with the type of JIA. These findings may allow the use of carbonyls as clinical markers of inflammatory process activity in patients with different types of JIA. It is also a potential tool for monitoring oxidative protein damage in other diseases and therapies.     

Immunotoxicological investigation of subacute combined exposure by permethrin and the heavy metals arsenic(III) and mercury(II) in rats

Effects of combined 28 days of oral exposure to the insecticide Permethrin (Pe), alone or in combination with arsenic-III (As) or Hg-II (Hg), were investigated on certain toxicological (body weight, organ weights), haematological (white blood cell (WBC) and red blood cell (RBC) counts, haematocrit (Ht), mean cell volume (MCV), cell content of the femoral bone marrow) and immune function (IgM-PFC, delayed-type hypersensitivity (DTH) reaction) parameters of male Wistar rats. Immunotoxic (H = high) and NOEL (L = low) doses of the three substances were determined in preliminary experiments under identical experimental conditions. In the present study, the immunotoxic dose of Pe (126 mg/kg) was combined with the NOEL dose of As (3.33 mg/kg) or Hg (0.40 mg/kg), and the NOEL dose of Pe (12.6 mg/kg) with the immunotoxic dose of As (13.3 mg/kg) or Hg (3.20 mg/kg). A separate group of animals, treated with the appropriate high dose component only, was used as internal control. Significant interactions were observed in the liver weight of the animals treated with Pe(H)-As(L) or As(H)-Pe(L), in the cell content of the femoral bone marrow in case of Pe(H)-As(L) and Pe(H)-Hg(L) combinations, as well as in the number of PFCs formed from 10(6) spleen cells in the Pe(H)-As(L) and in the maximum of DTH reaction in the Hg(H)-Pe(L) combination. The results show that combined exposures by the investigated substances modify the toxic (including immunotoxic) effects of the single compounds. These findings rise the probability that the interactions observed can also be present in human situations altering the health hazard of this three chemicals.     

In vitro immunomodulatory activity of flavonoid glycosides from Urtica dioica L

The major compounds isolated from the methanolic extract of the aerial parts of Urtica dioica L. were determined as quercetin-3-O-rutinoside (1). kaempherol-3-O-rutinoside (2). and isorhamnetin-3-O-glucoside (3). by chromatographic, chemical (acidic hydrolysis) and spectral (UV, IR, (1)H-NMR, (13)C-NMR) methods. Their immunomodulatory activities were studied in vitro by chemotaxis (Boyden Migration Chamber) and intracellular killing activity (NBT reduction) tests. Compounds 1, 2, 3 and the total flavonoid fraction were determined to have significant chemotactic effects in 4, 8, 16 microg doses. According to the results of the NBT reduction test, all flavonoid glycosides showed high intracellular killing activity. The results of both assays confirmed the immunostimulatory activity of the flavonoid fraction and the isolated flavonoid glycosides on neutrophils suggesting that they could possibly be useful for treating patients suffering from neutrophil function deficiency and chronic granulomatous diseases.     

Detection of the effects of repeated dose combined propoxur and heavy metal exposure by measurement of certain toxicological, haematological and immune function parameters in rats

In the present study, an immunotoxicity test system, containing general toxicological (body weight gain, organ weights), haematological (WBC,RBC, Ht, mean cell volume of the RBCs, cell content of the femoral bone marrow), and immune function (PFC assay, DTH reaction) investigations, was used for detection the effects of a 4 weeks repeated low dose combined oral exposure of male Wistar rats with propoxur and the heavy metals arsenic or mercury. Two doses of the compounds were used: a higher one (the lowest dose which resulted in significant change of at least one parameter examined in previous dose-effect experiments), and a lower one (the highest dose which proved to be non-effective). The applied doses were: 8.51 and 0.851 mg kg(-1) of propoxur, 13.3 and 3.33 mg kg(-1) of NaAsO(2), and 3.20 and 0.40 mg kg(-1) of HgCl(2). In the combination treatment, the high dose of propoxur was combined with the low dose of arsenic or mercury, and the high doses of each heavy metals were combined with the low dose of propoxur. The main finding of this study was that some of the combinations significantly altered the relative weight of liver, adrenals and kidneys, related to both the untreated and the high dose internal control. Among the immune functions examined, only the PFC content of the spleen showed a trend of changes in certain combinations versus the corresponding high dose control. According to the present results, combined exposure with propoxur and the heavy metals examined can modify the detection limit of the single compounds and/or may alter their toxic effects.