Melamine exposure during the weaning period negatively affects ovarian reserve.

In this study, it was aimed to investigate the effects of melamine exposure since the weaning period on ovarian tissue and ovarian reserve. Melamine is illegally added to milk and formula to provide high false protein positivity. Female rats (the weaning period = 21 days old, n = 18) were divided into 3 groups. 0.1 mL saline was applied to the control group by gavage for 21 days. Fifty mg/kg and seventy-five mg/kg melamine was dissolved in 0.1 mL of saline and applied by gavage for 21 days, respectively. At the end of the experiment, plasma anti-Mullerian hormone (AMH) was measured, follicle count and ovarian diameter measurement were performed in the right ovaries, and flow cytometric analysis for apoptosis was performed in the left ovaries. While a statistically significant decrease was not observed in the number of the follicle and ovarian diameter between the control and melamine-treated groups (p > 0.05), a significant decrease in the corpus luteum and a significant increase in the number of atretic follicles were observed (p < 0.05). Apoptosis (Annexin V) increased in both melamine groups and AMH plasma level decreased significantly in the 75 mg/kg group (p < 0.05). Melamine exposure from the weaning (early postnatal) period may cause a decrease in ovarian reserve in parallel with a dose increase.

Methotrexate-induced hepatotoxicity in rats and the therapeutic properties of vitamin E: a histopathologic and flowcytometric research.

Aim of the study: Methotrexate (MTX) causes oxidative stress-related liver damage. Our objective was to investigate the protective effects of vitamin E against MTX-induced hepatotoxicity through histopathological methods and flow cytometry. Material and methods: The rats were assigned to four groups: Control (2 ml saline for 5 days), MTX (20 mg/kg intraperitoneally (i.p.) only on the initial day of the study), MTX + vitamin E (20 mg/kg MTX (i.p.) only on the first day, and 100 mg/kg vitamin E (i.p.) was applied for 5 days during the study), Vitamin E (100 mg/kg of vitamin E (i.p.) was given for five days). Histopathologic changes and the flow cytometric apoptotic index were evaluated for liver tissue. The Kruskal-Wallis test was used for comparisons between groups. The statistical significance level was accepted as p < 0.05. Results: In the histopathological analysis, hepatocyte degeneration, dilatation of sinusoids, mononuclear cell infiltration, hydropic degeneration in hepatocytes, vacuolization, and pycnotic nucleus were observed in the MTX group. In the MTX + vitamin E group, hepatocyte degeneration, pycnotic nuclei, and dilatation in sinusoids were significantly lower compared to the MTX group. In the MTX group, glycogen accumulation in hepatocytes was lower compared to the control group. In the MTX + vitamin E group, glycogen accumulation in hepatocy-tes was higher compared to the MTX group. The flowcytometric apoptotic index (AI) percentage in the MTX group was 34.4% and in the MTX + vitamin E group the value was 9.4%. Conclusions: Our results demonstrated that vitamin E ameliorates MTX-induced liver damage. Co-using vitamin E and MTX drugs will be beneficial for the treatment of various diseases.

Hepatotoxic effects of melamine exposure from the weaning period in rats: a flow cytometric, electron microscopic, and histopathologic study.

This study aims to investigate the effects of melamine exposure from the weaning period (21st postnatal days in rats) on liver tissue. Female Wistar albino rats (n = 18) were divided into three groups. About 0.1-ml saline was applied to the control group by gavage for 21 days from the postnatal 21st day. The second group was taken 50-mg/kg melamine (in 0.1-ml saline) and the third group was taken 75-mg/kg melamine (in 0.1-ml saline) p.o. On the postnatal 45th day, all rats were sacrificed under anesthesia. Then, liver tissues were cut into three parts and two of them placed in neutral formalin for histopathological and flow cytometric analysis, and one of them placed in 2.5% glutaraldehyde. Histopathological analysis was performed with hematoxylin & eosin, Masson trichrome, periodic acid Schiff stained sections, and also with transmission electron microscopy. Apoptosis (Annexin V positivity) was analyzed by flow cytometry. According to histopathological analysis, hepatocyte damage, sinusoidal dilatation, and inflammatory cell infiltration significantly increased in both melamine groups compared with the control group. Apoptosis significantly increased in the 50 and 75-mg melamine groups compared with the control group. In the results of transmission electron microscopy analysis, there was abnormal chromatin distribution in the hepatocyte nuclei, loss in the cristae of the mitochondria, and organelle loss in large areas in the cytoplasm in both melamine exposure groups. As result, melamine exposure from the weaning period causes liver damage with increasing doses.

Long-term effects of elevated intra-abdominal pressure on testes an experimental model of laparoscopy.

PURPOSE: To determine the long-term outcomes of pneumoperitoneum on the testes in an experimental laparoscopy model. MATERIALS AND METHODS: Twenty-four rats were divided into three groups: Group A, the control group; Group B, exposed to a 10 mmHg intra-abdominal pressure (IAP); and Group C, exposed to a 20 mmHg IAP with CO2 pneumoperitoneum for 60 minutes. After 6 weeks, the testes were removed, and testicular injury score and Johnson score were examined histologically. Germ cell apoptosis was also detected using flow cytometry. RESULTS: A significant difference was determined between all groups in terms of testicular injury scores, Johnson scores, and germ cell apoptosis percentages. For the testicular injury score, there were significant differences between the groups for the right testis (group A versus B, P = .009; group A versus C, P < .0001; and group B versus C, P = .001) and for the left testis (group A versus B, P = .001; group A versus C, P < .0001; and group B versus C, P = .002). Significant differences were determined in the Johnson scores for the right testis between all groups (group A versus B, P= .001; group A versus C, P < .0001; and group B versus C, P = .008, respectively). Percentage of apoptotic testis cells were significantly differed between all groups (P = .001 for each). CONCLUSION: This study shows that increased IAP during pneumoperitoneum causes histopathology and apoptotically-evident damage to the testes in the long-term, depending on the magnitude of IAP increase, which may cause sub/infertility. Considering the experimental nature of this study,further clinical studies are needed for a more decisive conclusion.

Circulating platelet-leukocyte aggregates in patients with inflammatory bowel disease.

BACKGROUND: Inflammatory bowel diseases (IBDs), Crohn's disease, and ulcerative colitis are considered to be chronic inflammatory disorders implicated with recurrent tissue damage to the intestine. There is a positive correlation between platelet-leukocyte aggregates and ischemic vascular risk. There are limited data about the relationship between platelet-leukocyte aggregates and IBD. This study was designed to determine whether platelet-leukocyte aggregates increase in IBD, and whether a relationship exists between the elevation of platelet-leukocyte aggregates and disease activity. METHODS: A total of 20 patients with IBD (16 with ulcerative colitis and 4 with Crohn's disease) and 20 healthy controls participated in our study. Nine patients were in active-phase IBD, whereas 11 patients were in inactive phase. To show the presence of thrombocyte aggregates, the monoclonal antibodies such as Isotype IgG1 mouse antihuman CD42b-PE (phycoerythrin) (Beckman Coulter IMI417), Isotype IgG1 mouse antihuman CD45-FITC (fluorescein isothiocyanate) (Beckman Coulter IM0782), and Isotype IgG2a mouse antihuman CD45RO-FITC (Beckman Coulter IMI247) were used. Additionally, the values of platelet-neutrophil aggregates were measured in peripheral blood samples using flow cytometry techniques. RESULTS: The levels of platelet-leukocyte aggregates in blood samples were found to be significantly higher during both the active and inactive phases in patients with IBD. There were no statistically significant differences between active-phase and inactive-phase patients. CONCLUSION: We determined that the patient group had significantly higher platelet-leukocyte aggregate levels compared with the control group. This finding suggests that platelet-leukocyte aggregates may play a role in the development of IBD.

In vitro determination of apoptotic effect of heparin on lymphoblasts by using flow cytometric DNA analysis and measurements of caspase-9 activation and cytochrome C level.

Heparin induces apoptosis on peripheral neutrophils, mononuclear cells of the healthy controls, and on lymphoblasts of the patients with acute lymphoblastic leukemia, in vitro. We studied the caspase-9 activity and cytochrome C level as the indicators of the apoptotic effect of heparin on lymphoblasts by the intrinsic pathway of apoptosis. Twenty samples of the patients with acute lymphoblastic leukemia were included in the study. Cytochrome C level and caspase-9 activity were concomitantly determined with the percentage of apoptotic lymphoblasts when incubated in 0, 10, and 20 U/mL heparin concentrations at 0, 1, and 2 hours. The percentages of apoptosis of lymphoblasts at the first hour were higher than those at 0 and 2 hours in 10 and 20 U/mL heparin concentrations, separately (P<0.05). The mean percentage of apoptosis of lymphoblasts in 20 U/mL heparin levels was significantly higher than those in 0 and 10 U/mL heparin levels at 1 and 2 hours (P<0.05). The highest apoptotic effect of heparin on lymphoblasts was determined at the first hour in 20 U/mL heparin concentration. The mean caspase-9 activitity at the first hour was significantly higher than the values at 0 and 2 hours in 10 and 20 U/mL heparin levels, separately (P<0.05). The mean caspase-9 activity in 20 U/mL heparin concentration was significantly higher than values in 0 and 10 U/mL heparin concentrations at 1 and 2 hours (P<0.05). The highest caspase-9 activity was determined in 20 U/mL heparin levels at the first hour. The mean cytochrome C level at the first hour was significantly higher than those at 0 and 2 hours in 10 and 20 U/mL heparin concentrations, separately (P<0.05). The highest cytochrome C level was determined in 20 U/mL heparin concentration at the first hour. We claimed that heparin induces the apoptosis of lymphoblasts by the activation of the intrinsic pathway.

Apopitotic effects of dopamine and dobutamine on neutrophils of premature neonates.

OBJECTIVE: The apopitotic effects of dopamine and dobutamine used for the treatment of hypotensive premature newborns on neonatal neutrophils were investigated. METHODS: Fifteen premature neonates, gestational age less than 34 weeks and birth weight less than 2500 g, were enrolled into the study. Neutrophils were isolated from the blood samples obtained for routine completed blood count, in the first 72nd hour of life. Neutrophil samples were divided into three tubes as dopamine, dobutamine, and control groups. Dopamine and dobutamine were added at 10(-4) M concentration into two separate tubes as the study groups (dopamine and dobutamine groups) and 0.9 % NaCl was added to the third tube as the control group. Apoptotic neutrophil percentages were measured in the three groups initially (hour 0) and at the first, second, fourth, and sixth hours by flow cytometric analysis. RESULTS: The mean percentages of neutrophil apoptosis that measured at the beginning was similar in three groups. It was significantly higher in dopamine and dobutamine groups than the control group at the first, second, fourth, and sixth hours (p < 0.0005). It was also significantly higher in dopamine group, than dobutamine group at the first, second, fourth, and sixth hours (p < 0.0005). CONCLUSIONS: Dopamine and dobutamine may have an apoptotic effect on the neutrophils of premature. As neutrophils are numerically and functionally immature in premature infants, have a high risk for infection, these drugs should be used carefully and dobutamine may be preferred if there is no reason for special preference.

The effect of heparin on the cell cycle in human B-lymphoblasts: An in vitro study.

OBJECTIVE: Heparin has been shown to be a strong inhibitor of the proliferation of several cell types. In this in vitro study, we investigated whether different heparin concentrations can affect the cell cycle of lymphoblasts in newly diagnosed acute lymphoblastic leukemia (ALL) patients. METHODS: Lymphoblasts were incubated in different heparin concentrations (0, 10, 20 U/ml), and the percentages of lymphoblasts in each phase of the cell cycle were simultaneously measured by flow cytometry at 0, 1, and 2 hours (h). RESULTS: The percentages of lymphoblasts at the G2/M and S phases were significantly increased in 20 U/ml heparin concentration at 1 h compared to 0 U/ml (without heparin) concentration. We demonstrated that heparin increases the percentages of lymphoblasts in the S and G2/M phases in a concentration- and time-dependent manner. CONCLUSION: It was shown that heparin expands the proliferation of lymphoblasts by increasing the transition to G2/M and S phases and the S-phase fraction ratio. Heparin thus appears promising for its contribution to new treatment fields such as by providing a synergistic effect with chemotherapeutic drugs.

In vitro investigation of the apoptotic effect of heparin on lymphoblasts by using flow cytometric DNA analysis and fluorometric caspase-3 and -8 activities.

The apoptotic effect of heparin on the lymphoblasts obtained from 12 newly diagnosed children with acute lymphoblastic leukemia (ALL) was investigated in vitro. The lymphoblasts were incubated with 0, 10, and 20 U/mL heparin concentrations at 0, 1, and 2 h. The percentages of the apoptotic lymphoblasts were calculated by flow cytometry (FCM), and activities of caspase-3 and -8 were simultaneously measured by fluorometric protease activity method. The apoptotic effect of heparin on the lymphoblasts was determined in 10 and 20 U/mL heparin concentrations (p < 0.005 and p < 0.001, respectively) while no apoptosis was detected in 0 U/mL heparin concentration at 0, 1, and 2 h. The apoptotic percentages of the lymphoblasts were higher at the first hour than those at 0 and 2 h in 10 and 20 U/mL heparin levels (p < 0.001). The highest apoptosis was found at first hour in 20 U/mL heparin concentration. Increased concentrations of heparin had an increasing effect on the percentages of the apoptotic lymphoblasts. Significantly higher caspase-3 and -8 activities were determined in 10 and 20 U/mL heparin concentrations than those in 0 U/mL heparin concentration at 0, 1, and 2 h (p < 0.001). There were no significant differences between the caspase-3 and -8 activities in 10 and 20 U/mL heparin concentrations at 1 and 2 h (p > 0.05), while statistically significant differences were simultaneously detected in the apoptotic rates of the lymphoblasts (p < 0.001). This may be due to that the study included the limited patients, or measurement of the caspase activities is a more sensitive method than the FCM analysis for determination of apoptosis because the activation time of the caspases takes a long time period. It was concluded that the apoptotic effect of heparin in vitro on lymphoblasts developed due to the extrinsic pathway of apoptosis via the caspase-3 and -8 activations in newly diagnosed ALL patients.

The effects of pentoxifylline on liver regeneration after portal vein ligation in rats.

AIM: To determine the effects of pentoxifylline, a methyl xanthine derivative on hepatic cell production of uninterferred lobe after portal vein branch ligation. METHODS: Sixty-six rats were randomly allocated into 9 groups with 8 rats in PVL groups and 6 rats in sham operation groups. The portal branches of the median and the lateral liver lobes, corresponding to approximately 70% of the liver volume were ligated in the PVL groups. The control group received 0.9% NaCl solution. The rats in the treatment groups received pentoxifylline at the dose of 50 mg/kg/dy. After 1, 2, 4 days of portal vein ligation in both PVL and PVNL lobes the levels of adenine nucleotides were determined and flowcytometric analysis of cell cycles were performed. RESULTS: On the first day of portal branch ligation energy charge was significantly lower, in pentoxifylline treated group comparing to pentoxifylline untreated group, both in PVL and PVNL lobes (P<0.05). Proliferative indexes were 0.38 and 0.29 in pentoxifylline treated and pentoxifylline untreated PVNL lobes respectively (P<0.05). CONCLUSION: Pentoxifylline treatment resulted in an increase of percentage of calls entering mitosis phase on the first day after PVL, somehow accelerating the regeneration process.

Flow cytometrical analysis of adhesion molecules, T-lymphocyte subpopulations and inflammatory markers in pterygium.

BACKGROUND/AIM: Pterygium is a relatively frequent ocular surface disease with an unexplained etiopathogenesis. Our study was carried out with the aim to identify the presence of inflammatory cells and mediators such as T-lymphocyte subgroups (CD4 and CD8), intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and human leukocyte antigen-DR (HLA-DR) in pterygium tissue. METHODS: Pterygium tissue, obtained from 24 patients, and normal conjunctival tissue, from the nasal bulbar conjunctiva obtained from 14 patients operated for ocular perforations or vitrectomy, were separated into epithelial and stromal components under the microscope and suspended with phosphate-buffered saline solution to form a suspension. Cell suspensions were treated with specific antibodies for ICAM-1, VCAM-1, and HLA-DR and T-lymphocyte subgroups and evaluated with flow cytometry. The obtained data were compared statistically. RESULTS: When compared to the control tissue samples, higher rates of ICAM-1-positive cells, VCAM-1-positive cells and HLA-DR-positive cells were recorded in pterygium tissue samples. CD4 and CD8 lymphocytes were also found to be at higher levels when compared to the control group. There was a statistically significant difference between the two groups. CONCLUSION: When compared with normal conjunctival tissue, pterygium tissue had increased levels of T-lymphocyte infiltration and inflammatory markers demonstrating the possible contribution of cellular immunity to the pathogenesis.

Comparision of the apoptotic effects on lymphoblasts and on increase of myeloid lineage cells of a short-time, high-dose methylprednisolone and the conventional-dose prednisolone treatments in children with acute lymphoblastic leukemia.

The authors compare the apoptotic effect on the lymphoblasts and the proliferative effect on the myeloid lineage cells of a short-course high-dose methylprednisolone (HDMP) and the conventional-dose prednisolone treatments in children with acute lymphoblastic leukemia (ALL). The patients were divided into 2 groups. Group I (n = 10) received HDMP (30 mg/kg/day for 7 days) in a single dose before 6 a.m. perorally. Group II (n = 10) received prednisolone (2 mg/kg/day for 7 days) in 3 doses. The apoptotic percentages of lymphpblasts and the percentages of blasts and myeloid lineage cells were determined after performing the bone marrow aspiration (BMA) at diagnosis on the 0th, 3rd, and 7th days of the treatments in all patients. The mean apoptotic percentages of the lymphoblasts on the 3rd day were significantly higher than those on the 0th and 7th days in both groups (p < .05). The highest apoptosis was determined on the 3rd day in group I. The mean percentages of the blast cells on the 7th day were significantly lower than those on the 0th and the 3rd days in both groups (p < .05). The lowest lymphoblast percentage was determined on the 7th day in group I. The mean percentages of the CD13+ and CD33+ cells on the 7th day were significantly higher than those on the 0th and the 3rd days in both groups (p < .05). The highest percentages of the CD13+ and CD33+ cells were found on the 7th day in group I. Prednisolone and HDMP showed no proliferative effect on the CD14+ cells. These findings indicate that a short-course HDMP treatment shows a more effective apoptosis on the lymphoblasts and on the increase of the myeloid lineage cells when compared to the prednisolone treatment. The authors suggest that HDMP may be used in the treatment of patients with ALL instead of prednisolone.

Apoptotic and morphological features of the umbilical artery endothelium in mild and severe pre-eclampsia.

BACKGROUND: The pathology of the umbilical arterial endothelium in normal pregnancy and in pregnancy complicated with pre-eclampsia remains unclear. In this study the changes that occur in the umbilical artery endothelial cells were examined and endothelial cell morphology and apoptosis were compared among control, mild, and severe pre-eclamptic subjects. METHODS: Umbilical cords with a gestational age of between 35 and 40 weeks were collected from women with normal pregnancies (n=17), mild pre-eclampsia (n=10), and severe pre-eclampsia (n=12). We studied the umbilical artery endothelial cells using flow cytometry, and light and electron microscopy. Apoptosis was measured using flow cytometry and the terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling technique. The Kruskall-Wallis variance analysis and Mann-Whitney U-tests as post hoc were applied. RESULTS: In mild pre-eclamptics, the endothelial cells appeared ultrastructurally separated. A dilated endoplasmic reticulum, swollen mitochondria, and vanished mitochondrial cristae were observed. In severe pre-eclamptics, the cells were disorganized, highly contracted and vacuolated, separated from each other, and protruding prominently into the lumen. The percentages of endothelial cells that underwent apoptosis in mild (p<0.017) and severe pre-eclamptics (p<0.017) were higher than those in the controls. These apoptosis values were highest in severe pre-eclamptics (p<0.0001). CONCLUSION: Apoptosis and structural disruptions in the arterial endothelium of severe pre-eclamptics were prominent in all subjects. Increased endothelial apoptosis and structural disruptions are clinically related to intensity of pre-eclampsia, and may be associated with adaptation of the endothelial cells to pre-eclampsia.

Ham's F-10 medium and Ham's F-10 medium plus vitamin E have protective effect against oxidative stress in human semen.

OBJECTIVES: To investigate possible protective effects of vitamin E and Ham's F-10 medium (HF-10) on lipid peroxidation, apoptosis, motility, and vitality of spermatozoa. METHODS: Normozoospermic semen samples were obtained from 35 volunteers. Normal saline solution, HF-10 only, or HF-10 with vitamin E were added to split semen samples (control, group 1, and group 2, respectively). Sperm motility and vitality were evaluated at the end of 1, 2, and 24 hours. Superoxide dismutase, catalase, and malondialdehyde levels were assessed at the end of the first hour. Flow cytometric DNA analysis was performed at the end of 24 hours. RESULTS: Superoxide dismutase, sperm motility, and vitality were not different among the groups. The catalase values decreased in group 1, but not in group 2. Malondialdehyde values in supernatants decreased in group 2 and apoptosis of spermatozoa decreased in groups 1 and 2. CONCLUSIONS: Our data suggest that vitamin E and HF-10 protect against the reactive oxygen species-mediated damage on spermatozoa.

Therapeutic use of limbal stem cells.

Stem cells are defined as relatively undifferentiated cells that have the capacity to generate more differentiated daughter cells. Limbal stem cells are responsible for epithelial tissue repair and regeneration throughout the life. Limbal stem cells have been localized to the Palisades of Vogt in the limbal region. Limbal stem cells have a higher proliferative potential compared to the cells of peripheral and central cornea. Limbal stem cells have the capacity to maintain normal corneal homeostasis. However, in some pathological states, such as chemical and thermal burns, Stevens-Johnson syndrome, and ocular pemphigoid limbal stem cells fail to maintain the corneal epithelial integrity. In such situations, limbal stem cell transplantation has been required as a therapeutic option. In unilateral disorders, the usual source of stem cells is the contralateral eyes, but if the disease is bilateral stem cell allografts have to be dissected from family members or cadaver eyes. The advent of ex vivo expansion of limbal stem cells from a small biopsy specimen has reduced the risk of limbal deficiency in the donor eye. Concomitant immunosuppressive therapy promotes donor-derived epithelial cell viability, but some evidences suggest that donor-derived epithelial stem cell viability is not sustained indefinitely. Thus, long-term follow-up studies are required to ascertain whether donor limbal stem cell survival or promotion of recolonization by resident recipient stem cells occurs in restored recipient epithelium. However, this is not an easy task since a definitive limbal stem cell marker has not been identified yet. This review will discuss the therapeutic usage of limbal stem cells in the corneal epithelial disorders.

Comparison of the effects of ketotifen fumarate 0.025% and olopatadine HCl 0.1% ophthalmic solutions in seasonal allergic conjunctivities: a 30-day, randomized, double-masked, artificial tear substitute-controlled trial.

BACKGROUND: Topical antiallergic agents, such as antihistamines and mast-cell stabilizers, are the main therapeutic options for seasonal allergic conjunctivitis (SAC). Ketotifen fumarate and olopatadine HCl have dual action that offers a combination of these 2 mechanisms. Although clinical studies comparing the efficacy of these 2 drugs have shown that both were effective in the treatment of SAC, the results were contradictory and did not include the effects of these drugs on inflammatory markers. OBJECTIVES: The aims of this study were to compare the clinical efficacy of topical ketotifen and olopatadine eye drops and to determine the effects of these 2 drugs on the expression of cell adhesion molecules (CAMs) and inflammatory markers in conjunctival surface cells in patients with SAC. METHODS: This 30-day, randomized, double-masked, artificial tear substitute (ATS)-controlled clinical trial was conducted at the Department of Ophthalmology, Karadeniz Technical University, School of Medicine, Trabzon, Turkey. Patients with SAC were included in the study and randomly assigned to 1 of 3 groups: topical ketotifen fumarate 0.025% ophthalmic solution, topical olopatadine HCl 0.1% ophthalmic solution, or ATS (control group). All drugs were administered 2 drops per eye BID for 30 days. At the beginning of the study (day 0; baseline), on day 15, and on day 30, clinical scores (itching, tearing, redness, eyelid, swelling, and chemosis) and conjunctival impression cytology specimens were obtained. The percentages of cells expressing intercellular adhesion molecule 1, vascular CAM-1, human leukocyte antigen-DR, and beta1-integrin (CD29) from conjunctival impression cytology specimens were determined using flow cytometry. Patients were questioned about adverse events (AEs) at each visit. Ocular discomfort on installation of the drugs was recorded as an AE. RESULTS: Thirty-nine patients (20 men, 19 women; age range, 18-61 years) with SAC were included. Twelve patients received ketotifen; 13, olopatadine; and 14, ATS. In both active-treatment groups, the improvements of clinical scores (tearing and itching) were more pronounced compared with those in the ATS group, although the day-30 difference in tearing score between the olopatadine and ATS groups was not statistically significant. No significant within-group or between-group differences in mean scores for redness, chemosis, or eyelid swelling were found. The expression rates of CAMs and inflammatory markers in conjunctival surface cells were significantly more reduced with ketotifen and olopatadine compared with ATS. However, clinical and flow cytometric parameters were improved with ATS at 15 and 30 days compared with baseline. No AEs were observed during the study period. CONCLUSIONS: In this short-term study in a selected, small study population with SAC, ketotifen and olopatadine diminished the expression of CAMs and inflammatory markers on the conjunctival surface cells effectively. Both active treatments were more efficacious compared with ATS and were well tolerated.

Effects of Turkish pollen and propolis extracts on respiratory burst for K-562 cell lines.

Bee-collected pollen and propolis are apicultural products which are composed of nutritionally valuable substances and contain considerable amounts of polyphenol substances which may act as potent antioxidants. We wanted to show if respiratory burst within a cancer cell lines could be influenced when incubated with pollen and propolis extracts or not. Pollen and propolis extracts at concentrations of 50, 25, 12.5 and 0 mg/ml were prepared by dimethyl sulfoxide (DMSO). K-562 cell cultures and mononuclear cell (MNC) cultures prepared from a peripheral blood sample to serve as control cells were incubated with extracts for 24 h. Determination of respiratory burst was carried out by intracellular dichlorofluorescein (DCFH) test by using flow-cytometric fluorescence analysis. While about 90% and 66% fluorescence was detected at zero concentrations for both K-562 and MNC cultures, fluorescence positivity decreased (between 3.8% and 11.8%) as concentrations of both propolis and pollen extracts increased for K-562 cell culture, but unchanged (between 20% and 83%) for MNC culture. It was concluded that pollen and propolis extracts inhibit respiratory burst within cancer cell lines probably by their antioxidant potentials.

Platelet function disorder in patients with coronary slow flow.

BACKGROUND: Some patients evaluated for chest pain with angiographically normal coronary arteries show coronary slow flow phenomenon (CSFP) on angiography. Slow flow of dye in epicardial coronary arteries is also not an infrequent finding in patients during routine coronary angiography. The precise pathophysiology of CSFP is not known yet. HYPOTHESIS: This study investigates the presence of platelet function disorders in patients with CSFP. METHODS: The patient group included 24 patients with CSFP detected by coronary angiography via the TIMI "frame count" method, and a control group included 23 patients with normal coronary flow. Platelet aggregability induced by use of ristocetin, collagen, and adenosine diphosphate (ADP), was measured from all blood samples in both control and patient groups. RESULTS: The ratio of platelet aggregability increased significantly in patients with CSFP compared with patients with normal coronary flow (ristocetin 57.6 +/- 15 vs. 45.4 +/- 17.1, collagen 62.9 +/- 16.4 vs. 48.9 +/- 25.3, ADP 59.4 +/- 18 vs. 42.4 +/- 15.2, p < 0.05). CONCLUSION: Platelet aggregability is increased in patients with CSFP.

Effects of dexamethasone, all-trans retinoic acid, vitamin D(3) and interferon-alpha on FO myeloma cells.

BACKGROUND: Since multiple myeloma responds poorly to conventional chemotherapy or radiotherapy, new therapeutic approaches are needed. This study investigated the effects of dexamethasone, all-trans retinoic acid (ATRA), the active metabolite of vitamin D(3) [1,25(OH)(2)D(3)] and interferon-alpha on FO mouse myeloma cells (non-immunoglobulin-secreting myeloma cell line) in single drug or drug combination groups in vitro. METHODS: Apoptosis ratio and change in cell counts in 4 single drug groups (dexamethasone, ATRA, vitamin D(3) and interferon-alpha) and 6 combination drug groups (dexamethasone + vitamin D(3,) dexamethasone + ATRA, dexamethasone + interferon-alpha, vitamin D(3) + ATRA, vitamin D(3) + interferon-alpha, interferon-alpha + ATRA) were compared with the control group. RESULTS: When treatment groups were compared with the control group, there was a significant increase in apoptosis in all, but this was most prominent in the group treated with dexamethasone alone. The apoptosis ratios were 0.10 and 6.82% in the control and dexamethasone-only groups, respectively. We also found that there was a significant decrease in cell count, particularly in the dexamethasone-only, ATRA-only, and ATRA-vitamin D(3) combination groups. CONCLUSION: ATRA, interferon-alpha, vitaminD(3) and particularly dexamethasone have significant effects on FO mouse myeloma cells resulting in a decreased cell count and an increased apoptosis ratio. This study should be repeated with human myeloma cell lines for further information.

In vitro determination of the apoptotic effect of heparin on lymphoblasts using DNA analysis and measurements of Fas and Bcl-2 proteins by flow cytometry.

Heparin has an apoptotic effect beside its anticoagulant, anti-inflammatory, antihypertensive, and antiproliferative effects. In this study, the authors detected the percentages of apoptotic lymphoblasts and the expressions of apoptotic Fas protein and antiapoptotic Bcl-2 protein with flow cytometry in vitro after the incubation of lymphoblasts with heparin. Eleven newly diagnosed acute lymphoblastic leukemia (ALL) children were included in the study. Lymphoblasts were incubated in all different levels of heparin concentrations (0, 10, and 20 U/mL) and the percentages of apoptotic lymphoblasts and the percentages of Fas protein and Bcl-2 proteins were simultaneously measured by flow cytometry at 0, 1, and 2 h. At 0, 1, and 2 h, apoptosis was determined when heparin was added in 10- and 20-U/mL concentrations (p <.05). The apoptotic effect of heparin on lymphoblasts was higher at the first hour than at 0 and 2 h in 10- and 20-U/mL heparin concentrations (p <.01). The highest apoptosis was detected in the 20-U/mL heparin concentration at the first hour. The expression levels of Fas protein on lymphoblasts were higher at the first hour than at 0 and 2 h in 10- and 20-U/mL heparin concentrations (p <.001). The highest expression of Fas protein was observed in the 20-U/mL heparin concentration at the first hour. The expression levels of Bcl-2 protein on lymphoblasts were lower at the first hour than at 0 and 2 h in 10- and 20-U/mL heparin concentrations (p <.001). The lowest expression of Bcl-2 protein was detected in the 20-U/mL heparin concentration at the first hour. Increased concentrations of heparin had an increasing effect on the percentages of apoptotic lymphoblasts. The expression percentages of Fas protein on lymphoblasts also increased, whereas the expression percentages of Bcl-2 protein on lymphoblasts decreased (p <.05). These results suggest that low-dose heparin may cause significant apoptosis of lymphoblasts in newly diagnosed ALL patients.