Sándor Szűcs

Phagocytic index of neutrophil granulocytes and monocytes in healthy and preeclamptic pregnancy

Neutrophil granulocytes and monocytes have been intensively studied, but there is no scientific data on one of their most important functions, namely the phagocyte function in pregnancy and preeclampsia. The aim of this study was to examine this function. Twenty-five healthy pregnant, 25 preeclamptic pregnant, and 20 healthy, non-pregnant women were enrolled into our study. Cells were isolated from peripheral blood samples, marked and evaluated for the phagocytic index with an immunofluorescent microscope after phagocytosing the zymosan molecules. The phagocytic function of monocytes and neutrophil granulocytes decreased significantly in healthy pregnancy compared with non-pregnant women and in preeclampsia, and it decreased significantly compared with healthy pregnancy. Decreased phagocytic function in healthy pregnancy can be a part of the maternal immunosuppression, which is essential for the protection of the hemiallograft fetus. Further reduction of phagocytic function may be one of the immunoregulatory abnormalities found in preeclampsia.

Aliphatic alcohol contaminants of illegally produced spirits inhibit phagocytosis by human granulocytes

Abstract Context: Unregulated production of spirits in many countries leads to products containing appreciable levels of aliphatic alcohols (AAs) and is the main source of human exposure to these substances worldwide. Previous studies have confirmed that alcohol abuse can lead to ethanol-induced immunosuppression and thereby increased susceptibility to infectious diseases. Granulocytes, as professional phagocytic cells, play a crucial role in engulfment and killing of pathogenic microorganisms. Thus, a decrease in their phagocytic activity has been invoked as a factor in the impaired antimicrobial defense observed in alcoholics. However, AAs consumed as contaminants of illicit spirits may also influence phagocytosis, thereby contributing to a further decrease in microbicidal activity but, so far, this has not been studied. Objective: Therefore, the aim of this study was to measure granulocyte phagocytosis following treatment of granulocytes with those higher alcohols found in illegal spirits. Materials and methods: Granulocytes were isolated from human peripheral blood. Then phagocytosis of opsonized zymosan particles by granulocytes treated with AAs individually and in combination was determined. Results: These alcohols inhibited phagocytosis in a concentration-dependent manner and at lower concentrations when combined than when tested individually. Discussion and conclusion: Due to their synergistic effects, it is possible that, in combination with ethanol, they may inhibit phagocytosis in a clinically meaningful way in episodic heavy drinkers.

Susceptibility of lung epithelial cells to alkylating genotoxic insult

Alkylation is one of the most common types of DNA damage that can lead to mutations and cancer. Lung is the primary target organ of airborne alkylators such as ethylene oxide (EO). However, the ability of EO to cause lung cancer has not been clearly demonstrated yet. The aim of this study was to investigate the susceptibility of lung cells to alkylating DNA insult by detecting EO‐mediated DNA damage with the alkaline comet assay in human lung epithelial cells, peripheral blood lymphocytes, and keratinocytes. The susceptibility of these cell types toward the alkylating insult induced by EO was compared against the oxidative DNA insult induced by hydrogen peroxide (H2O2). Due to the volatility of EO, its active concentrations were monitored by gas chromatography during exposure and were found to decrease significantly in a time‐dependent manner. EO induced a statistically significant genotoxic effect at the lowest concentration used (16.4 µM) in lung epithelial cells and in lymphocytes, while in keratinocytes, a genotoxic effect was not detected until 55.5 µM EO. However, lung epithelial cells demonstrated increased resistance to oxidative insult. In fact, oxidative DNA damage detectable by endonuclease treatment was minimal in lung cells compared with the other cell types. These results suggest an increased sensitivity of lung epithelial cells toward the alkylating effects of EO, which was not observed for oxidative DNA damage. Our findings point out the importance of DNA alkylation and the possible role of EO on the induction of lung cancer. Environ. Mol. Mutagen. 54:682–689, 2013.

Aliphatic alcohols of illegally produced spirits can act synergistically on superoxide-anion production by human granulocytes.

Context: Aliphatic alcohols present in illegally produced spirits in a large number of low and middle income countries have been implicated in the etiology of chronic liver disease and cirrhosis. Previous studies have confirmed that chronic alcoholism can lead to increased susceptibility to infectious diseases. Reduced superoxide-anion (O2·−) production by granulocytes could provide a mechanism by which antimicrobial defense is impaired in alcoholics. In vitro experiments have also demonstrated that ethanol can inhibit granulocyte O2·− generation. Aliphatic alcohols consumed as contaminants of illicit spirits may also influence O2·− production thereby contributing to a decrease in microbicidal activity. Objective: The aim of this study was to investigate this possibility. It measured the O2·− production by human granulocytes following treatment of the cells with aliphatic alcohol contaminants found in illicit spirits. Materials and methods: Granulocytes were isolated from human buffy coats with centrifugal elutriation and then treated with individual aliphatic alcohols and their mixture. The O2·− production was stimulated with phorbol-12-13-dibutyrate and N-formyl-methionyl-leucyl-phenylalanine (FMLP) and measured by superoxide dismutase inhibitable reduction of ferricytochrome c. Results: Aliphatic alcohols of illegally produced spirits inhibited the FMLP-induced O2·− production in a concentration dependent manner. They suppressed O2·− generation at 2.5–40 times lower concentrations when combined than when tested individually. Discussion and conclusion: Aliphatic alcohols found in illegally produced spirits can inhibit FMLP-induced O2·− production by granulocytes in a concentration-dependent manner. Due to their synergistic effects, it is possible that, in combination with ethanol, they may inhibit O2·− formation in heavy episodic drinkers.

Granulocyte superoxide anion production and regulation by plasma factors in normal and preeclamptic pregnancy

Data on the respiratory burst activity of granulocytes from healthy and preeclamptic women have remained contradictory. To investigate the role of reactive oxygen species in the etiology of preeclampsia we measured superoxide anion generation by granulocytes from non-pregnant, healthy, and preeclamptic women. We also examined the reciprocal effects of heat-inactivated and non-inactivated plasma on superoxide production. Superoxide generation was measured by ferricytochrome-c reduction. Superoxide production induced by either phorbol-12,13-dibutirate or N-formyl-methionyl-leucyl-phenylalanine was significantly decreased in granulocytes from normal pregnant women compared with non-pregnant and preeclamptic women. The phorbol-12,13-dibutirate-induced superoxide generation by granulocytes from non-pregnant and preeclamptic women was significantly inhibited by plasma from healthy pregnant women. The N-formyl-methionyl-leucyl-phenylalanine-stimulated superoxide production by granulocytes from non-pregnant and preeclamptic women was suppressed only by non-inactivated plasma, not heat-inactivated plasma from healthy pregnant women. Plasma from preeclamptic women did not influence the phorbol-12,13-dibutirate- and N-formyl-methionyl-leucyl-phenylalanine-induced superoxide production by control granulocytes. The phorbol-12,13-dibutirate-induced superoxide generation by granulocytes from healthy pregnant women was significantly increased by the effect of plasma from non-pregnant and preeclamptic women, but when stimulating with N-formyl-methionyl-leucyl-phenylalanine only non-inactivated plasma caused the same enhancement. These data indicate that reduced superoxide generation in normal pregnancy may be caused by maternal immunosuppressive factors present in plasma. The failure to reduce superoxide production in preeclampsia may be partly responsible for the endothelial dysfunction characteristic of that condition.

Aliphatic alcohols in spirits inhibit phagocytosis by human monocytes

Abstract A large volume of alcoholic beverages containing aliphatic alcohols is consumed worldwide. Previous studies have confirmed the presence of ethanol-induced immunosuppression in heavy drinkers, thereby increasing susceptibility to infectious diseases. However, the aliphatic alcohols contained in alcoholic beverages might also impair immune cell function, thereby contributing to a further decrease in microbicidal activity. Previous research has shown that aliphatic alcohols inhibit phagocytosis by granulocytes but their effect on human monocytes has not been studied. This is important as they play a crucial role in engulfment and killing of pathogenic microorganisms and a decrease in their phagocytic activity could lead to impaired antimicrobial defence in heavy drinkers. The aim of this study was to measure monocyte phagocytosis following their treatment with those aliphatic alcohols detected in alcoholic beverages. Monocytes were separated from human peripheral blood and phagocytosis of opsonized zymosan particles by monocytes treated with ethanol and aliphatic alcohols individually and in combination was determined. It was shown that these alcohols could suppress the phagocytic activity of monocytes in a concentration-dependent manner and when combined with ethanol, they caused a further decrease in phagocytosis. Due to their additive effects, it is possible that they may inhibit phagocytosis in a clinically meaningful way in alcoholics and episodic heavy drinkers thereby contribute to their increased susceptibility to infectious diseases. However, further research is needed to address this question.

Effect of normal and preeclamptic plasma on superoxide-anion production of neutrophils from healthy non-pregnant women

This study has examined whether production of superoxide-anion by granulocytes differs between non-pregnant, healthy pregnant and preeclamptic pregnant women. First, we assessed superoxide-anion production in 13 non-pregnant women, 11 healthy pregnant women and 14 preeclamptic pregnant women. Then, we examined the effect of plasma samples of healthy pregnant and preeclamptic pregnant women on superoxide production by neutrophils separated from healthy pregnant women. Superoxide generation was measured by ferricytochrome-c reduction. Phorbol-12,13-dibutyrate- and n-formyl-methionyl-leucyl-phenylalanine-stimulated superoxide-anion production was significantly decreased in healthy pregnant womens granulocytes compared with non-pregnant women. There was no significant difference between granulocyte superoxide-anion production in preeclamptic pregnant and non-pregnant women. When neutrophils from non-pregnant women were incubated in plasma from healthy pregnant women, the granulocyte phorbol-12,13-dibutyrate-stimulated superoxide-anion production was significantly inhibited. With the same stimulator, there were no significant differences between superoxide-anion production of neutrophils incubated in autologous, non-pregnant and preeclamptic pregnant plasma. If n-formyl-methionyl-leucyl-phenylalanine was used for stimulation, there were no significant differences in the superoxide-anion production of granulocytes in either group. Granulocyte superoxide-anion production decreases during pregnancy; this decrease does not occur in preeclampsia, and may cause endothelial damage. It is conceivable that there are unidentified factors in maternal circulation which inhibit superoxide-anion production by granulocytes in healthy pregnant women.

The effect of healthy pregnant plasma and preeclamptic plasma on the phagocytosis index of neutrophil granulocytes and monocytes of nonpregnant women

ABSTRACT Objective: Phagocyte function of neutrophil granulocytes and monocytes is decreased in healthy pregnancy and further decreased in preeclampsia. The cause of the declined function is unknown. Decreased phagocyte function can lead to the higher infection rate in healthy pregnancy and may also be responsible for the increased susceptibility to infections and high trophoblast concentration in preeclampsia. The aim of this study is to examine the phagocyte function of neutrophil granulocytes and monocytes. Methods: Monocytes and neutrophil granulocytes were separated from the peripheral circulation of six nonpregnant patients and incubated in plasma samples from six healthy pregnant, six preeclamptic pregnant, and six nonpregnant patients. The cells were marked and evaluated for the phagocytosis index with immunofluorescent microscope after phagocyting the zymosan molecules. Results: Phagocyte function of neutrophils as well as monocytes from nonpregnant patients were decreased significantly when the cells were incubated in plasma samples from healthy pregnant patients, and further decreased when incubated in plasma samples from preeclamptic pregnant women. Conclusion: The decreased phagocyte function of neutrophil granulocytes and that of monocytes in healthy pregnancy and the further decreased phagocyte function in preeclampsia is caused by factor(s) in the maternal circulation.

Consumption of home-made spirits is one of the main source of exposure to higher alcohols and there may be a link to immunotoxicity

We are grateful to Dr. Lachenmeier and his colleague for their interest in our article. We were surprised by his comments but wonder whether they reflect a difference in terminology in that we included aliphatic alcohols in illegally produced spirits within the category of ‘‘contaminants’’. Dr. Lachenmeier claim that 1-propanol, 1-butanol and isobutanol are considered as ‘‘flavouring agents’’ when spirits contain these substances at certain concentrations. However, those manufacturing spirits seek to avoid their presence at high levels, while methanol, also a component of some spirits, is not a ‘‘flavouring agent’’. We believe that, regardless of the choice of terminology, it is important to know whether the presence of these substances in alcoholic beverages has an impact on human immune system, so justifying our investigation. In our previous work, we found that aliphatic alcohols were more frequently present in home-made spirits than in commercial spirits. We detected methanol in 82% (13% in commercial spirits), 2-butanol in 94% (55% in commercial spirits), 1-propanol 100% (81% in commercial spirits), isobutanol in 100% (81% in commercial spirits), 1-butanol 47% (6% in commercial spirits) and isoamyl alcohol in 100% (71% in commercial spirits) of the home-made spirit samples. Furthermore, the concentration of aliphatic alcohols was significantly higher in home-made spirits than in those of from commercial sources. This is consistent with Dr. Lachenmeier’s own research where he also found higher alcohols more common in unrecorded products (samohon for personal use or for sale) than in commercially manufactured alcohol (vodka). Where we differ is our view about the relative amount of legally and illegally produced spirits being consumed, but this is not something that can be resolved definitively with the data available. If it is felt necessary, the meaning of our ‘‘critical sentence’’ could be easily resolved by modifying it as follows: ‘‘Consumption of contaminated unrecorded alcohols, particularly illicitly produced spirits is one of the most common forms of exposure to AAs affecting large populations worldwide’’. However, we do rather feel that we are getting into semantics. The statement that ‘‘the concentration of higher alcohols in unrecorded alcohols is significantly lower than in commercial alcohol’’ can be disputed for the following reasons. The average level of higher alcohols in unrecorded products shown in their Figure 1 (presented in Lachenmeier D W, Rehm J. What is the main source of human exposure to higher alcohols and is there a link to immunotoxicity? Immunopharmacol Immunotoxicol 2013;35:451–453) is derived from previous studies in which different types of unrecorded alcoholic products were analyzed. These included home-made spirits, illegally imported and untaxed alcohols, smuggled alcoholic beverages, counterfeit or relabeled alcohols, surrogate alcohols, medicinal alcohols, wines and beers from several European countries. However, these studies were not representative and suffer from two important limitations. First, the collection of samples was not systematic and, except in Poland, Russia and Ukraine, only unrecorded products were obtained and examined but their commercially produced, and thus recorded, counterparts were not compared. Second, although to the World Health Organization’s definition of unrecorded alcohol encompasses all of the products tested, this is actually a very heterogeneous group, being produced from different raw materials (grain, a variety of fruits and grapes), by different technologies, including fermentation and brewing (beer), fermentation (wine), fermentation and distillation (spirits). In addition, some smuggled alcohols are industrially produced, subject to quality control and recorded in their countries of origin. *These authors share the first authorship.

Monocyták és neutrophil granulocyták fagocitafunkciója ovariumcarcinomában

INTRODUCTION Recently, tumor-infiltrating immune cells have been studied in various cancers. However, fewer studies address the role of peripheral immune cells in the pathogenesis of cancer. AIM Our aim was to investigate whether the phagocytic activity of peripheral monocytes and neutrophil granulocytes is affected by the removal of tumor in advanced ovarian cancer. METHOD We investigated peripheral blood samples from 12 patients with advanced stage of serous epithelial ovarian cancer - which were collected before the optimal tumor reduction surgery and on the 7th postoperative day - and from 8 healthy women. After separation of monocytes and neutrophils, the cells were incubated with opsonized fluorescein isothiocyanate-labeled zymosan A particles as the target of phagocytosis. By using fluorescence microscope we counted the number of particles phagocytized by the cells and calculated the phagocytic index. Statistical analysis of the data was performed using analysis of variances method. RESULTS Preoperative phagocytic indexes of monocytes and neutrophils from patients were significantly lower than phagocytic indexes of the corresponding cells from healthy women. The phagocytic function of monocytes and granulocytes isolated from postoperative samples of patients significantly increased compared to preoperative values and reached the phagocytic indexes of monocytes and neutrophils from healthy controls. CONCLUSION Based on our results we assume that the tumor and/or its microenvironment in ovarian cancer may produce factors that can depress the phagocytic function of monocytes and granulocytes. Since the phagocytic indexes increased following the cytoreductive surgery, it can be assumed that after the removal of the tumor, the production of these factors is reduced or eliminated. Orv Hetil. 2018; 159(33): 1353-1359.