Histone acetylation and arachidonic acid cytotoxicity in HepG2 cells overexpressing CYP2E1.

The aim of this work was to assess the role of ethanol-derived acetate and acetate-mediated histone acetylation in arachidonic acid-induced stress in HepG2 cells and cells overexpressing CYP2E1. Cells were grown for 7 days with 1 mM sodium acetate or 100 mM ethanol; their acetylated histone proteins and histone deacetylase 2 expression was quantified using Western blot. Ethanol- or acetate-pretreated cells were also treated for 24 h with 60 µM arachidonic acid to induce oxidative stress. Cytotoxicity was estimated by lactate dehydrogenase release, 3-[4,5-dimethylthiazolyl-2] 2,5-diphenyltetrazolium bromide test, and by DNA damage, while oxidative stress was quantified using dichlorofluorescein diacetate. Cells grown with ethanol or acetate had increased acetylated histone H3 levels in both cell types and elevated acetylated histone H4 levels in cells overexpressing CYP2E1 but not in naïve cells. In cells overexpressing CYP2E1 grown with ethanol, expression of histone deacetylase 2 was reduced by about 40 %. Arachidonic acid altered cell proliferation and was cytotoxic mostly to cells engineered to overexpress CYP2E1 but both effects were significantly lower in cells pretreated with ethanol or acetate. Cytotoxicity was also significantly decreased by 4-methylpyrazole--a CYP2E1 inhibitor and by trichostatin--an inhibitor of histone deacetylases. In cells pretreated with acetate or ethanol, the oxidative stress induced by arachidonic acid was also significantly lower. Our data indicate that histone hyperacetylation may in some extent protect the cells against oxidative stress. It is possible that acetate may act as an antioxidant at histone level. This mechanism may be relevant to alcohol-induced liver injury.

The mentally ill: the way they perceive their own illness and their expectations from the society.

People diagnosed as mentally ill face the suffering of their own disease as well as the social stigmatization, which, in turn, can aggravate their psychopathological symptoms and exacerbate their social isolation. Mental diseases and their symptoms cause an ill person to seem recondite for the people around. She or he feels different from everybody else, thus requires special care, sensitivity, and respect from the others. A supportive and full of acceptation environment is indispensable to optimize socio-professional therapy and rehabilitation of the ill. The society should possess some knowledge of mental diseases and respect the dignity and human rights of the ill. The aim of our work was to show how the mentally ill perceive their illness as well as specify the patients' expectations concerning their relations with the healthy people from their environment. This work consists of a theoretical part, a research which contains the results of a questionnaire, and conclusions. There were 52 patients who took part in the research. They were all aware of their illness, although the research was held during the lighter phase of the disease or its remission. The analysis of the collected data indicates that the ill perceived their disease as a burden and obstacle to fulfilling their principal aims. The disease also engendered negative emotional reactions in them and significantly impaired the quality of their life.

Attitude toward alcoholics and their families.

The objective of this work was to find out whether alcoholics undergoing therapy are considered rightful members of society. We also examined the attitudes of society toward the alcoholic's family. Alcoholism is a specific disorder. Alcohol-addicted persons deny their illness and often engage their closest surrounding in the denial system. They also find in the society a specific margin of consent to drink. Research shows that treated alcoholics believe the society has a low level of knowledge concerning alcohol addiction. They fear the perspective of going back to work or finding a new job after finishing therapy. Studies done among the inhabitants of the mountainous Podhale Region in Poland show that a high percentage of people are convinced that one should be ashamed of being an alcoholic and many would feel awkward in the presence of a sober alcoholic. The people declare that they would not trust a person being treated because of the addiction and name professions that alcoholics should not work in. The spouse of an alcoholic is often blamed for the addiction, and children of alcoholics are perceived as people with worse life perspectives than their peers.

Cladribine combined with cyclophosphamide and mitoxantrone as front-line therapy in chronic lymphocytic leukemia.

The objective of the study was to determine the effectiveness and the toxicity of a combined chemotherapy consisting of cladribine (2-CdA), mitoxantrone and cyclophosphamide (CMC regimen) in the treatment of previously untreated B cell chronic lymphocytic leukemia (B-CLL). From August 1998 to December 2000 2-CdA was administered at a dosage of 0.12 mg/kg for 3 (CMC3) or 5 (CMC5) consecutive days, mitoxantrone at 10 mg/m2 on day 1 and cyclophosphamide at 650 mg/m2 on day 1 to 62 patients with advanced or progressive B-CLL. The cycles were repeated at 4 week intervals or longer if severe myelosuppression occurred. Twenty patients received CMC5 and 42 patients CMC3. Within the analyzed group an overall response (OR) rate (CR+PR) of 64.5% (95% CI: 52.7-76.3%) was reported, including 29.0% CR. There was no difference in the CR rate between the patients treated with CMC5 (30%) and CMC3 (28.6%) (P = 0.9), nor in the OR rate (55.0% and 69.0%, respectively, P = 0.3). Residual disease was identified in seven out of 18 (38.9%) patients who were in CR, including two treated with CMC5 and five treated with CMC3 protocols. CMC-induced grade III or IV thrombocytopenia occurred in 12 (19.4%) of patients, including four (20%) CMC5-treated and eight (19%) CMC3-treated patients (P= 0.8). Neutropenia grade III or IV was observed in seven (35%) and 11 (26.2%) patients, respectively (P = 0.8). Severe infections, including pneumonia and sepsis, occurred more frequently after CMC5 (11 patients, 55.0%) than CMC3 (10 patients, 28.6%) (P = 0.03) Fourteen patients died, including six treated with CMC5 and eight treated with CMC3 (30% and 19%, respectively). Infections were the cause of death in nine patients, including four in the CMC5 group and five in the CMC3 group. In conclusion, our results indicate that the CMC programme is an active combined regimen in previously untreated B-CLL patients; its efficiency seems to be similar to that observed earlier in B-CLL patients treated with 2-CdA as a single agent. However, toxicity, especially after CMC5 administration, is significant. Therefore, we recommend the CMC3 but not the CMC5 programme for further evaluation.