scholarly journals Effect of Staphylococcus aureus and Streptococcus uberis on apoptosis of bovine mammary gland neutrophils in vitro

2012 ◽  
Vol 50 (No. 1) ◽  
pp. 11-23 ◽  
Author(s):  
Z. Sladek ◽  
D. Rysanek ◽  
M. Faldyna
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Death ◽  
Mammary Gland ◽  
Programmed Cell Death ◽  
Propidium Iodide ◽  
Annexin V ◽  
Dna Fragments ◽  
Bovine Mammary Gland ◽  
Streptococcus Uberis

Neutrophils play an important role in the defence of the bovine mammary gland against bacterial infections. In the course of the resolution of mammary gland inflammation, neutrophils undergo programmed cell death – apoptosis. The aim of this study was to confirm whether the co-cultivation of neutrophils of the bovine mammary gland with either Staphylococcus aureus or Streptococcus uberis leads to signs of apoptosis. In the study, 16 mammary glands of four virgin heifers aged 16 to 18 months were examined. Neutrophils were obtained by lavage after an induced influx. After a three-hour incubation of the neutrophils with bacteria in vitro, neutrophil apoptosis was detected by morphological features, by determination of histone-associated DNA fragments (ELISA), and by Annexin -V and propidium iodide positivity (flow cytometry). S. aureus and S. uberis reduced the incidence of karyopycnotic and zeiotic neutrophils (P < 0.01), and insignificantly reduced the concentration of histone -associated DNA fragments (P > 0.05). The incubation of neutrophils with bacteria, however, increased the proportion of Annexin –V-positive cells (P < 0.01) and Annexin -V and propidium iodide-positive cells (P < 0.05). Co-cultivation of neutrophils with either S. aureus or S. uberis led to the induction of phosphatidylserine translocation characteristic of the early stage of apoptosis. The late signs of apoptosis were delayed by co-cultivation of neutrophils with both pathogens. Therefore it is obvious that although the programmed cell death of apoptosis is initiated by these pathogens, the completion of the program is delayed.

Type II secretory phospholipase A2 binds to ischemic flip-flopped cardiomyocytes and subsequently induces cell death

2003 ◽  
Vol 285 (5) ◽  
pp. H2218-H2224 ◽  
Author(s):  
R. Nijmeijer ◽  
M. Willemsen ◽  
C. J. L. M. Meijer ◽  
C. A. Visser ◽  
R. H. Verheijen ◽  
...  
Keyword(s):  
Cell Death ◽  
Propidium Iodide ◽  
Caspase 3 ◽  
Ischemia Reperfusion ◽  
Annexin V ◽  
Border Zone ◽  
Metabolic Inhibitors ◽  
Type Ii ◽  
Secretory Phospholipase

Type II secretory phospholipase A2 (sPLA2) is a cardiovascular risk factor. We recently found depositions of sPLA2 in the necrotic center of infarcted human myocardium and normally appearing cardiomyocytes adjacent to the border zone. The consequences of binding of sPLA2 to ischemic cardiomyocytes are not known. To explore a potential effect of sPLA2 on ischemic cardiomyocytes at a cellular level we used an in vitro model. The cardiomyocyte cell line H9c2 or adult cardiomyocytes were isolated from rabbits that were incubated with sPLA2 in the presence of metabolic inhibitors to mimic ischemia-reperfusion conditions. Cell viability was established with the use of annexin V and propidium iodide or 7-aminoactinomycin D. Metabolic inhibition induced an increase of the number of flip-flopped cells, including a population that did not stain with propidium iodide and that was caspase-3 negative. sPLA2 bound to the flip-flopped cells, including those negative for caspase-3. sPLA2 binding induced cell death in these latter cells. In addition, sPLA2 potentiated the binding of C-reactive protein (CRP) to these cells. We conclude that by binding to flip-flopped cardiomyocytes, including those that are caspase-3 negative and presumably reversibly injured, sPLA2 may induce cell death and tag these cells with CRP.

Phagocytosis of Staphylococcus aureus by bovine mammary gland macrophages and intracellular protection from antibiotic action in vitro and in vivo

1984 ◽  
Vol 51 (4) ◽  
pp. 513-523 ◽  
Author(s):  
Neil Craven ◽  
James C. Anderson
Keyword(s):  
Staphylococcus Aureus ◽  
Mammary Gland ◽  
Antibiotic Therapy ◽  
Bovine Mammary Gland ◽  
Lactating Cows ◽  
Colony Forming Units ◽  
Subsequent Exposure ◽  
Increased Sensitivity

SummaryMacrophages isolated from the involuted bovine mammary gland were cultured in vitro. Phagocytosis of opsonized Staphylococcus aureus occurred rapidly, but intracellular killing of bacteria was slow. Many intracellular staphylococci survived for up to 4 d exposure to extracellular cloxacillin and emerged from within the macrophages to multiply extracellularly when the antibiotic was inactivated. Rifampicin was significantly more efficient than cloxacillin in killing intracellular S. aureus after 18 h incubation, but it too failed to sterilize the cultures within 3 d. Staphylococci, which had remained viable within macrophages during 20 h incubation with extracellular cloxacillin, showed an increased sensitivity to dilute lysostaphin on subsequent exposure. A 3 d course of intramammary therapy with cloxacillin, commencing simultaneously with an infecting inoculum of ∼108 colony forming units (c.f.u.) S. aureus, apparently eliminated the infection from one quarter of the udders of each of three lactating cows, but bacteria were re-isolated from two cows after a delay of several days. However, when other quarters of the same cows were infected with ∼108 c.f.u. S. aureus which had been phagocytosed by autologous mammary macrophages, similar simultaneous antibiotic therapy failed to affect these infections. The in vitro and in vivo findings indicate the significance of intracellular survival of S. aureus as a factor contributing to failure of antibiotic therapy.

scholarly journals The effect of the bacterial pathogens Staphylococcus aureus and Streptococcus uberis on morphological features of apoptosis of heifers mammary gland neutrophils

2005 ◽  
Vol 53 (4) ◽  
pp. 61-74 ◽  
Author(s):  
Tereza Langrová ◽  
Zbyšek Sládek ◽  
Dušan Ryšánek
Keyword(s):  
Staphylococcus Aureus ◽  
Mammary Gland ◽  
Morphological Changes ◽  
Bacterial Pathogens ◽  
Light And Electron Microscopy ◽  
Neutrophil Granulocytes ◽  
Apoptotic Cells ◽  
Streptococcus Uberis ◽  
Qualitative Changes

The aim of the student thesis was to review the effect of the significant bacterial pathogens,Staphylococcus aureusand Steptococcus uberis on programmed cell death – apoptosisin vitro, and to determine the dynamic of morphological changes during aging neutrophil granulocytes of heifers mammary glandin vitro. Using light and electron microscopy we have noted characteristic alterations of apoptotic neutrophils. These are running in three consequential phases. We found out, that the interaction of bacterial pathogens with neutrophils during the incubation have lead in expressive quantitative and qualitative changes in apoptotic cells proportion. Concretely, the influence of both pathogens on mammary gland neutrophils caused the defer of apoptosis expression. Here,S. aureuscaused lower number of apoptotic neutrophils in comprasion withS. uberis. The outcomes testified, thatS. aureusandS. uberisinteraction with heifers mammary gland neutrophilsin vitrocauses alterations relating to apoptosis of these cells. Looking at the results of the study, we can conclude, that the pathogensS. aureusandS. uberisare not only significant heifers mammary gland – affection causers, but they significantly influence the cells of defensive system in their functions too. They significantly decrease the appereance of morphological apoptosis manifestations on neotrophils of tissue pool of the heifers mammary gland. The numbers of apoptotic cells in neutrophil population confirm, that during the interaction with mentioned pathogens the defer of morphological apoptosis manifestations happens. Then, higher number of apoptotic neutrophils in stages of apoptotic corpuscles implies increasing dynamic of this process. Beside that, the dynamic of apoptotic process is influenced by the specifity of certain bacterious actor too.

scholarly journals Action of inhibitors Heat shock proteins 90 and 27 on dexamethasone-induced apoptosis of tumor cells

2010 ◽  
Vol 9 (3) ◽  
pp. 68-71 ◽  
Author(s):  
Ye. V. Kaigorodova ◽  
N. V. Ryazantseva ◽  
V. V. Novitsky ◽  
A. N. Maroshkina ◽  
M. V. Belkina ◽  
...  
Keyword(s):  
Cell Death ◽  
Heat Shock ◽  
Programmed Cell Death ◽  
Heat Shock Protein ◽  
Tumor Cells ◽  
Propidium Iodide ◽  
Fluorescent Microscopy ◽  
Annexin V ◽  
Induced Apoptosis ◽  
Shock Proteins

Programmed cell death of tumor cells of line Jurkat in conditions of cultivation with various concentration of dexamethasone, selective inhibitors of Hsp90 (Heat shock protein — Hsp) (17-AAG) and Hsp27 (KRIBB3) was investigated. An estimation of realisation apoptosis spent by method of fluorescent microscopy with use annexin V-FITC and propidium iodide. Inhibition of Hsp90 and Hsp27 leads to activation of tumor cells Jurkat apoptotic program and strengthening of dexamethasone-induced apoptosis. Hsp27 and Hsр90 play an antiapoptotic role in tumor cells of line Jurkat.

scholarly journals In vitro antioxidant treatment recovers proliferative responses of anergic CD4+ lymphocytes from human immunodeficiency virus-infected individuals

Blood ◽  
1996 ◽  
Vol 87 (11) ◽  
pp. 4746-4753 ◽  
Author(s):  
A Cayota ◽  
F Vuillier ◽  
G Gonzalez ◽  
G Dighiero
Keyword(s):  
Human Immunodeficiency Virus ◽  
Cell Death ◽  
Programmed Cell Death ◽  
Redox Status ◽  
Therapeutic Strategies ◽  
Antioxidant Treatment ◽  
Immunodeficiency Virus ◽  
Cd4 Lymphocytes ◽  
Cellular Thiol

Oxidative stress has been proposed to be involved in the immunologic defeat observed in effector calls of the immune system as well as in lymphocyte cell death and viral replication in human immunodeficiency virus (HIV)-infected patients. Because thiol-containing antioxidants such as N-acetyl-L-cysteine have been shown to have beneficial effects on CD4+ lymphocyte survival and to inhibit programmed cell death and HIV-1 replication, they may play a role in therapeutic strategies of this disease. In this work we have studied the cellular thiol levels and the affect of in vitro antioxidant treatment of purified CD4+ lymphocytes from HIV-infected patients, and correlated these parameters to proliferative responses and programmed cell death. We show that CD4+ lymphocytes from HIV-infected patients display impaired proliferative responses and a significant decrease in cellular thiol levels, indicating a disturbed redox status. Interestingly, antioxidant treatment succeeded to restore defective proliferative responses to CD3- mediated activation in 8 of 11 patients (high antioxidant responders). In contrast to high responders, patients failing to respond to antioxidant treatment (low antioxidant responders), were characterized by an abnormal ratio of apoptotic cells, which was not affected by N- acetyl-L-cysteine and/or 2-beta-mercaptoethanol preincubation. These results demonstrate for the first time that antioxidant treatment is able to revert the impaired proliferative activity of CD4 cells from HIV-infected patients and could help designing therapeutic strategies with antioxidant drugs. However, this action is not observed in cells undergoing programmed cell death.

Histochemical localization and possible antibacterial role of xanthine oxidase in the bovine mammary gland

1988 ◽  
Vol 55 (1) ◽  
pp. 25-32 ◽  
Author(s):  
Robert A. Collins ◽  
Keith R. Parsons ◽  
Terry R. Field ◽  
A. John Bramley
Keyword(s):  
Antibacterial Activity ◽  
Mammary Gland ◽  
Xanthine Oxidase ◽  
Histochemical Localization ◽  
Bovine Mammary Gland ◽  
Streptococcus Uberis ◽  
Antibacterial Assay ◽  
Secretory Tissue ◽  
Teat Canal

SummaryXanthine oxidase (XO) was demonstrated to be present in the teat canal and secretory tissue of the bovine mammary gland by histochemical techniques. Homogenates of these tissues were able to replace XO in an antibacterial assay with Streptococcus uberis. The action of XO on its substrate hypoxanthine was shown to provide an essential component for anti-streptococcal activity mediated by lactoperoxidase. A mechanism is proposed whereby the interaction of XO, lactoperoxidase and thiocyanate may provide antibacterial activity in the teat canal.

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