Influence of adrenaline and ephedrine on primary DNA damage of lymfocytes of man in vitro

U ovom radu cilj istraživanja je bilo ispitivanje primarnih oštećenja DNK izolovanih limfocita čoveka pod uticajem adrenalina i efedrina. Oštećenja DNK su evaluirana primenom in vitro Komet testa. Ispitivan je širok spektar koncentracija adrenalina i efedrina (u rasponu od 0,0005 μM do 500 μM) u različitim vremenskim intervalima (15 min, 60 min, 120 min, 240 min i 24 sata). Najizraženije oštećenje DNK ustanovljeno je nakon 15 min tretmana adrenalinom, pri čemu su sve koncetracije izuzev najniže (0.0005 μM) dovele do povećane migracije DNK. Nakon 60 min, 120 min, 240 min tretmana adrenalinom indukovano je oštećenje DNK u opsegu od 5 do 300 μM. Najslabiji efekat ispoljen je nakon 24 sata, tako da su samo najviše koncetracije adrenalina (150 μM i 300 μM) dovele do povećanog stepena oštećenja DNK. Radi utvrđivanja mogućeg učešća reaktivnih kiseoničnih vrsta (ROS) u indukovanju DNK oštećenja pod dejstvom adrenalina upotrebili smo antiokisanse katalazu (100 IU i 500 IU) i kvercetin (100 μM i 500 μM). Kotretman limfocita adrenalinom (300 μM) i antioksidansima nakon 15 ili 60 minuta doveo je do značajnog smanjenja količine DNK u repovima kometa. Prema tome, može se zaključiti da adrenalin ispoljava svoje genotoksične efekte indukcijom reaktivnih kiseoničnih vrsta i da se neka oštećenja poprave tokom prva četiri sata nakon tretmana adrenalinom. Za razliku od adrenalina, efedrin nije doveo do povećanja migracije DNK u odnosu na negativnu kontrolu tokom različitih vremenskih intervala. Jedino je koncetracija efedrina od 500 μM nakon 15 minuta tretmana indukovala oštećenje DNK koje je bilo statistički značajno.The objectives of these investigations were to investigate primary DNA damage in isolated human lymphocytes exposed to adrenaline and ephedrine. DNA damage was evaluated by the in vitro Comet assay. A broad spectrum of adrenaline and ephedrine concentrations (range from 0.0005 μM to 300 μM) were examined in the Comet assay for various treatment times (15 min, 60 min, 120 min, 240 min and 24 h). The most profound DNA damage was observed after 15 min of adrenaline treatment, as all concentrations tested except the lowest one (0.0005 μM) caused an increase in DNA migration. After 1 h, 2 h and 4 h of treatment, adrenaline induced DNA damage at concentration range from 5 μM to 300 μM. The slightest DNA damage was observed after 24 h of adrenaline treatment, therefore only the highest concentrations of adrenaline (150 μM and 300 μM) caused increased level of DNA damage. In order to evaluate the potential contribution of reactive oxygen species (ROS)-induced DNA damage exposed to adrenaline, we used antoxidants catalase (100 IU and 500 IU) and quercetin (100 μM and 500 μM). Co-treatment of lymphocytes with adrenaline (300 μM) and antioxidants for 15 or 60 min, significantly reduced the quantity of DNA in the comet tails. Therefore, it can be concluded that adrenaline exhibits genotoxic effects mainly through induction of reactive oxygen species and that some of the DNA damage is repaired during the first four hours following the treatment with adrenalin

Febrile neutropenia

Neutrofilni granulociti ili neutrofili imaju vrlo važnu ulogu u obrani organizma od nastanka infekcije. Broj neutrofila čini 60 – 70 % bijele krvne loze u perifernoj krvi, odnosno leukocita čiji normalan broj iznosi oko 2,5 – 7,5x109/L (ovisno o referentnim vrijednostima pojedinog laboratorija). Vrlo je važno obratiti pozornost na smanjenje broja neutrofila u perifernoj krvi, odnosno neutropeniju, jer sniženje broja neutrofila u perifernoj krvi može pogodovati nastanku infekcija, poglavito onih bakterijskih. Neutropenija može biti posljedica djelovanja različitih čimbenika, a jedna od njih je i primjena citotoksične terapije u onkoloških i hematoloških bolesnika, te nastaje izravnim toksičnim djelovanjem citotoksične terapije na stanice granulocitopoeze s posljedičnim kočenjem razvoja neutrofilnih granulocita. U bolesnika u kojih se primjenjuje citotoksična terapija česta je pojavnost febrilne neutropenije. Ovaj poremećaj u broju neutrofila predstavlja ijatrogeni sindrom za koji je karakteristična povišena tjelesna temperatura (> 38,5 °C) i pad broja neutrofila u perifernoj krvi (< 0,5x109/L). Febrilna neutropenija jedno je od hitnih stanja u radu s onkološkim bolesnicima i samo brzim prepoznavanjem i energičnim antibiotskim liječenjem moguće je spriječiti nastanak infekcije, komplikacije infekcije i mogući posljedični letalni ishod infekcije.Neutrophils or neutrophilic granulocytes have an important role in human immune response and serve to fight foreign organisms and initial infection. Neutrophil count ranges around 2,5 – 7,5x109/L (varies between laboratories). They represent about 60-70 % of all white blood cells present in peripheral blood. State with decreased absolute neutrophil count is called neutropenia. It is very important not to overlook neutropenia as it can lead to infection, especially bacterial. Neutropenia can be caused by a variety of factors, one of these being chemotherapy-induced febrile neutropenia. This type of neutropenia is caused by a direct toxic effect of cytotoxic agents on granulocytopoietic cells which in turn block neutrophil development. Febrile neutropenia represents an iatrogenic syndrome, generally characterized by fever (>38,5°C) along with decreased absolute neutrophil count (<0,5x109/L). It occurs frequently in patients receiving cytotoxic therapy. Febrile neutropenia is a hematological emergency which requires rapid detection and vigorous treatment with broad-spectrum antibiotics in order to stop infections, their complications and possible lethal outcome

Febrile neutropenia

Neutrofilni granulociti ili neutrofili imaju vrlo važnu ulogu u obrani organizma od nastanka infekcije. Broj neutrofila čini 60 – 70 % bijele krvne loze u perifernoj krvi, odnosno leukocita čiji normalan broj iznosi oko 2,5 – 7,5x109/L (ovisno o referentnim vrijednostima pojedinog laboratorija). Vrlo je važno obratiti pozornost na smanjenje broja neutrofila u perifernoj krvi, odnosno neutropeniju, jer sniženje broja neutrofila u perifernoj krvi može pogodovati nastanku infekcija, poglavito onih bakterijskih. Neutropenija može biti posljedica djelovanja različitih čimbenika, a jedna od njih je i primjena citotoksične terapije u onkoloških i hematoloških bolesnika, te nastaje izravnim toksičnim djelovanjem citotoksične terapije na stanice granulocitopoeze s posljedičnim kočenjem razvoja neutrofilnih granulocita. U bolesnika u kojih se primjenjuje citotoksična terapija česta je pojavnost febrilne neutropenije. Ovaj poremećaj u broju neutrofila predstavlja ijatrogeni sindrom za koji je karakteristična povišena tjelesna temperatura (> 38,5 °C) i pad broja neutrofila u perifernoj krvi (< 0,5x109/L). Febrilna neutropenija jedno je od hitnih stanja u radu s onkološkim bolesnicima i samo brzim prepoznavanjem i energičnim antibiotskim liječenjem moguće je spriječiti nastanak infekcije, komplikacije infekcije i mogući posljedični letalni ishod infekcije.Neutrophils or neutrophilic granulocytes have an important role in human immune response and serve to fight foreign organisms and initial infection. Neutrophil count ranges around 2,5 – 7,5x109/L (varies between laboratories). They represent about 60-70 % of all white blood cells present in peripheral blood. State with decreased absolute neutrophil count is called neutropenia. It is very important not to overlook neutropenia as it can lead to infection, especially bacterial. Neutropenia can be caused by a variety of factors, one of these being chemotherapy-induced febrile neutropenia. This type of neutropenia is caused by a direct toxic effect of cytotoxic agents on granulocytopoietic cells which in turn block neutrophil development. Febrile neutropenia represents an iatrogenic syndrome, generally characterized by fever (>38,5°C) along with decreased absolute neutrophil count (<0,5x109/L). It occurs frequently in patients receiving cytotoxic therapy. Febrile neutropenia is a hematological emergency which requires rapid detection and vigorous treatment with broad-spectrum antibiotics in order to stop infections, their complications and possible lethal outcome

Febrile neutropenia

Neutrofilni granulociti ili neutrofili imaju vrlo važnu ulogu u obrani organizma od nastanka infekcije. Broj neutrofila čini 60 – 70 % bijele krvne loze u perifernoj krvi, odnosno leukocita čiji normalan broj iznosi oko 2,5 – 7,5x109/L (ovisno o referentnim vrijednostima pojedinog laboratorija). Vrlo je važno obratiti pozornost na smanjenje broja neutrofila u perifernoj krvi, odnosno neutropeniju, jer sniženje broja neutrofila u perifernoj krvi može pogodovati nastanku infekcija, poglavito onih bakterijskih. Neutropenija može biti posljedica djelovanja različitih čimbenika, a jedna od njih je i primjena citotoksične terapije u onkoloških i hematoloških bolesnika, te nastaje izravnim toksičnim djelovanjem citotoksične terapije na stanice granulocitopoeze s posljedičnim kočenjem razvoja neutrofilnih granulocita. U bolesnika u kojih se primjenjuje citotoksična terapija česta je pojavnost febrilne neutropenije. Ovaj poremećaj u broju neutrofila predstavlja ijatrogeni sindrom za koji je karakteristična povišena tjelesna temperatura (> 38,5 °C) i pad broja neutrofila u perifernoj krvi (< 0,5x109/L). Febrilna neutropenija jedno je od hitnih stanja u radu s onkološkim bolesnicima i samo brzim prepoznavanjem i energičnim antibiotskim liječenjem moguće je spriječiti nastanak infekcije, komplikacije infekcije i mogući posljedični letalni ishod infekcije.Neutrophils or neutrophilic granulocytes have an important role in human immune response and serve to fight foreign organisms and initial infection. Neutrophil count ranges around 2,5 – 7,5x109/L (varies between laboratories). They represent about 60-70 % of all white blood cells present in peripheral blood. State with decreased absolute neutrophil count is called neutropenia. It is very important not to overlook neutropenia as it can lead to infection, especially bacterial. Neutropenia can be caused by a variety of factors, one of these being chemotherapy-induced febrile neutropenia. This type of neutropenia is caused by a direct toxic effect of cytotoxic agents on granulocytopoietic cells which in turn block neutrophil development. Febrile neutropenia represents an iatrogenic syndrome, generally characterized by fever (>38,5°C) along with decreased absolute neutrophil count (<0,5x109/L). It occurs frequently in patients receiving cytotoxic therapy. Febrile neutropenia is a hematological emergency which requires rapid detection and vigorous treatment with broad-spectrum antibiotics in order to stop infections, their complications and possible lethal outcome

Selected hematology ratios in cats with non-septic effusions highly suspected of feline infectious peritonitis

In veterinary medicine, knowledge about hematologic ratios (neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR) and mean platelet volume-to-platelet ratio (MPV/PLT)) is limited, particularly in cats. While the roles of these ratios have been proven in oncology, systemic inflammation with or without systemic inflammatory response syndrome (SIRS), and sepsis, information is lacking about their alterations in non-septic effusions, like feline infectious peritonitis (FIP). This study aimed to describe whether NLR, PLR, and MPV/PLT were changed and whether they correlated with routine hematologic and biochemical parameters in 16 cats with non-septic effusions, highly suspected to be the effusive form of FIP without SIRS, compared to nine clinically healthy cats. The NLR was calculated as the absolute count of neutrophils divided by the absolute count of lymphocytes, PLR by calculating the absolute platelet divided by the absolute lymphocyte count, and MPV/PLT by dividing mean platelet volume by absolute platelet count. The NLR, MPV, and MPV/PLT ratios were higher in cats with non-septic effusions suspected to be FIP, but PLR did not differ, when compared to healthy cats. Correlation analysis did not show any association between the selected ratios and hematological and biochemical parameters. In the absence of leukocytosis, increased NLR could help us to confirm the presence of systemic inflammation in cats with non-septic effusions indicative of FIP. However, a high MPV/PLT ratio should be interpreted with caution, especially in cats

Some traits of Verotoxin-producing strains of Escherichia coli isolated from cattle

Verotoxin-producing Esherichia coli (VTEC) is one of six pathogenicity groups of Eschericha coli. The reservoir for VTEC is the intestinal tract of domestic animals, primarily ruminants. Investigations in our geographic region have also demonstrated that domestic animals are a significant VTEC reservoir. In spite of this, sporadic diseases in humans caused by these agents are rare, and no epidemics have been registered so far. The question is, therefore, what percentage of VTEC isolated in our region have the characteristics of enterohaemorrhagic E. coli (EHEC) which cause intestinal diseases in humans. The aim of this study was to test the isolated bovine strains for some phenotypic and genotypic traits and to determine the percent of strains that belong to EHEC. A total of 105 VTEC strains isolated from cattle were tested for the presence of verotoxin (vtx) genes by PCR (polymerase chain reaction) method. All of them possessed one or more vtx genes. In accordance with our investigations, only four (3.8%) strains, of a total of 105 VTEC strains belong to groups of EHEC. It may be concluded that the majority of strains isolated from cattle in this part of the world do not have phenotypic traits typical for EHEC. Therefore human VTEC associated diseases in Serbia are rare, despite the fact that domestic animals frequently harbor VTEC

Oxidative stress and DNA damage in peripheral blood mononuclear cells from normal, obese, prediabetic and diabetic persons exposed to adrenaline in vitro

Diabetes represents one of the major health concerns, especially In developed countries. Some hormones such as the stress hormone adrenaline can induce reactive oxygen species (ROS) and may worsen the diabetes. Therefore, the main aim of the investigation was to find out whether peripheral blood mononuclear cells (PBMCs) from normal persons have less DNA damage induced by adrenaline (0.1, 1 and 10 mu M) in comparison to PBMCs from obese, prediabetic and diabetic patients. Also, the biochemical parameters of oxidative stress (TBARS, catalase) and lactate dehydrogenase were monitored. It was observed that higher concentrations of adrenaline (1 and 10 mu M) induced DNA damage in the obese, prediabetic and diabetic groups. In healthy individuals only the highest concentration of adrenaline caused significant Increase in the DNA damage. In summary, total comet score (TCS) comparison has shown significant differences between groups, and DNA damaging effects of adrenaline were most evident in diabetic patients. The results of the biochemical analysis also demonstrate that adrenaline exerts most obvious effects in diabetic individuals which is manifested as significant change of parameters of oxidative stress. In summary, the obtained results demonstrated that diabetics are more sensitive to genotoxic effects of adrenaline and this effect probably resulted from decreased antioxidative defence mechanisms in various stages of progression through diabetes. Therefore, these results could contribute to a better understanding of a role of endocrine factors to damage of cellular biomolecules which could be useful in finding novel therapeutic approaches and lifestyle changes with an aim to lower the possibility of diabetes complications