Immunohistochemical investigation of P16, P53 and Ki-67's prognostic values in diffuse large B-Cell lymphomas.

AIM: The aim of this study is to determine the immunohistochemical properties of Ki-67, P53 expression and loss of P16, and to assess their relationship with both clinical parameters and patient survival in DLBCL. METHOD: Forty patients, diagnosed at the Pathology Department of our institute with nodal DLBCL were selected as the study group. The relationship between P16, P53, Ki-67 expressions and clinical and laboratory parameters like age, gender, performance status, Eastern Cooperative Oncology Group (ECOG), clinical stage, presence of B-symptoms, bone marrow involvement, International Prognostic Index (IPI) score, lactate dehydrogenase (LDH) level, extranodal extension, relapse, C-reactive protein (CRP), sedimentation, number of leukocytes in patients and patient survival were then statistically evaluated. RESULTS: Our results display no statistically significant correlation between P53 expression and loss of P16, Ki-67 proliferation index and clinical parameters and overall survival (p > 0.05). The only statistically significant relationship was between loss of P16 and stage (p 0.05). CONCLUSION: According to the results of our study, the loss of P16, P53 gene expression and Ki-67 proliferation index have no effect on life expectancy of patients with DLBCL (Tab. 2, Fig. 2, Ref. 29).

Increased micronucleus frequency in phytohaemagglutinin-stimulated blood cells of patients with vitiligo.

BACKGROUND: Vitiligo is a relatively common, acquired pigmentary disorder characterized by areas of depigmented skin resulting from loss of melanocytes in the epidermis. Although several hypotheses have been proposed for the aetiology and pathogenesis of vitiligo, the cause of vitiligo remains unclear. OBJECTIVE: To evaluate spontaneous micronucleus (MN) frequency using the cytokinesis block MN assay to determine damages at the DNA or chromosome level in phytohaemagglutinin (PHA)-stimulated blood cells of patients with vitiligo and healthy control subjects. METHODS: Peripheral blood samples were obtained and cultured from 21 patients with vitiligo (mean age: 21.48 +/- 9.78 years) and 21 age- and sex-matched healthy control subjects (mean age: 21.52 +/- 9.80 years). MN values were scored in binucleated cells obtained from whole-blood cultures of patients and control subjects. RESULTS: MN frequencies (mean +/- SD) in PHA-stimulated blood cells of patients with vitiligo and control subjects were 0.94 +/- 0.58 and 0.58 +/- 0.32, respectively. Compared with control subjects, MN frequencies of patients with vitiligo were found significantly higher than those of the control subjects (P = 0.012). CONCLUSION: Our results indicate unexpectedly some chromosomal/DNA damage in whole-blood cultures of patients with vitiligo. We do not know, however, if these chromosome/DNA instabilities observed in the cells of vitiligo patients resulted from the cause or from the consequences of the disorder.

Increased micronucleus, nucleoplasmic bridge, nuclear bud frequency and oxidative DNA damage associated with prolactin levels and pituitary adenoma diameters in patients with prolactinoma.

Prolactinoma is the most common pituitary tumor. Most pituitary tumors are benign, but they often are clinically significant. We investigated cytokinesis-block micronucleus cytome (CBMN cyt) assay parameters and oxidative DNA damage in patients with prolactinoma to assess the relations among age, prolactin level, pituitary adenoma diameter and 8-hydroxy-2'-deoxyguanosine (8-OHdG) level in patients with prolactinoma. We investigated 27 patients diagnosed with prolactinoma and 20 age- and sex-matched healthy controls. We measured CBMN cyt parameters and plasma 8-OHdG levels in peripheral blood lymphocytes of patients with prolactinoma and controls. The frequencies of micronucleus (MN), nucleoplasmic bridge, nuclear bud, apoptotic and necrotic cells, and plasma 8-OHdG levels in patients with prolactinoma were significantly greater than controls. MN frequency was correlated positively with age, prolactin levels and pituitary adenoma diameters in patients with prolactinoma. The increased chromosomal and oxidative DNA damage, and the positive correlation between MN frequency, prolactin levels and pituitary adenoma diameters may be associated with increased risk of cancer in patients with prolactinoma, because increased MN frequency is a predictor of cancer risk.

Determination of Gibberellic Acid (GA3)-Induced Oxidative Stress in a Model Organism Galleria mellonella L. (Lepidoptera: Pyralidae).

The plant growth regulator gibberellic acid (GA3) is known to negatively impact growth and development of insects. In this study, larvae of Galleria mellonella L. (Lepidoptera: Pyralidae) were fed a diet with varying dosages of GA3 to investigate how antioxidant enzymes are influenced. Activity levels in last instars reared in laboratory at 25 ± 2°C, 60 ± 5% relative humidity, and a photoperiod of 12:12 (L:D) h were measured for superoxide dismutase (SOD), glutathione S-transferase (GST), and catalase (CAT). Treatment with GA3 in diet resulted in a remarkable increase in the activities of both SOD and GST at lower GA3 doses (50-1,000 ppm) with respect to control and higher doses. The activity of CAT in the hemolymph of last instars significantly increased at all doses when compared with that in the hemolymph of untreated larvae. This trend in the increase of CAT was not dose-wise, except for the significant increases at 2,000 and 5,000 ppm when compared with that of untreated and all treated groups. Consequently, our results showed that GA3 is effective at activating the antioxidant defense system of insects as a source of free radical and can be toxic for larvae in a dose-dependent manner. Therefore, we suggest that the increase in the activity of GST, SOD, and CAT in larvae may indicate a physiological adaptability to compensate for GA3-induced stress.

Effects of gibberellic acid on hemocytes of Galleria mellonella L. (Lepidoptera: Pyralidae).

The impacts of different doses of the plant growth regulator gibberellic acid (GA(3)) in diet on the number of total and differential hemocytes, frequency of apoptotic, and necrotic hemocytes, mitotic indices, encapsulation, and melanization responses were investigated using the greater wax moth Galleria mellonella L. (Lepidoptera: Pyralidae) larvae. Total hemocyte counts increased in G. mellonella larvae at all treatment doses whereas GA(3) application had no effect on the number of different hemocyte types. The occurrence of apoptosis, necrosis and mitotic indices in GA(3) treated and untreated last instars were detected by acridine orange or ethidium bromide double staining by fluorescence microscopy. While the ratio of necrotic hemocytes increased at all GA(3) treatments, that of late apoptotic cells was only higher at doses >200 ppm when compared with untreated larvae. The percentage of mitotic index also increased at 5,000 ppm. Positively charged DEAE Sephadex A-25 beads were used for analysis of the levels of encapsulation and melanization in GA(3) treated G. mellonella larvae. At four and 24 h posttreatments with Sephadex A-25 bead injection, insects were dissected under a stereomicroscope. Encapsulation rates of larval hemocytes were dependent on the extent of encapsulation and time but not treatment groups. While the extent of melanization of hemocytes showed differences related to time, in general, a decrease was observed at all doses of GA(3) treated larvae at 24 h. We suggest that GA(3) treatment negatively affects hemocyte physiology and cell immune responses inducing cells to die by necrosis and apoptosis in G. mellonella larvae.

Micronucleus frequency in the oral mucosa and lymphocytes of patients with Behçet's disease.

Behçet's disease (BD) is a chronic, multisystemic, inflammatory disorder characterized mainly by recurrent oral and genital aphthous ulcerations and uveitis. Our study aimed to determine the genetic damage in patients with BD. The micronucleus (MN) frequency was counted in peripheral lymphocytes and exfoliated cells of the patients with BD. MN analysis was performed in peripheral lymphocytes of 30 patients with BD and in 20 healthy controls by the cytokinesis-block method, and on uncultured cells of the oral cavity in 10 patients and 9 healthy controls. We found significantly higher MN rates in lymphocytes of the patients than the control subjects (P = 0.000). There were no significant differences between the patients with or without treatment (P = 0.860). The MN frequency in exfoliated cells of the patients was higher than in those of healthy controls (P = 0.013), and there was no significant difference between the exfoliated cells of the treated and untreated patients (P = 0.201). Our results indicate that genetic damage may play a secondary but important part in the aetiology of BD and that treatment with colchicine does not induce MN.