Comparison of HIF-1α and survivin levels in patients with diabetes and retinopathy of varying severity.

PURPOSE: This study measured serum hypoxia--inducible factor-1 (HIF-1α) and survivin levels in patients with diabetes and investigated their association with the severity of retinopathy. METHODS: This study included 88 patients with type 2 diabetes mellitus who underwent routine eye examinations. Three groups were created. Group 1 consisted of patients without diabetic retinopathy. Group 2 included patients with non-proliferative diabetic retinopathy. Group 3 included patients with proliferative diabetic retinopathy. To measure serum HIF-1α and survivin levels, venous blood samples were collected from patients. RESULTS: The mean HIF-1α levels in groups 1, 2, and 3 were 17.30 ± 2.19, 17.79 ± 2.34, and 14.19 ± 2.94 pg/ml, respectively. Significant differences were detected between groups 1 and 3 (p=0.01) and between groups 2 and 3 (p=0.01). The mean survivin levels in groups 1, 2, and 3 were 42.65 ± 5.37, 54.92 ± 5.55, and 37.46 ± 8.09 pg/ml, respectively. A significant difference was only detected between groups 2 and 3 (p=0.002). CONCLUSION: The present study revealed that serum HIF-1α and survivin levels are increased in patients with non-proliferative diabetic retinopathy compared to those in patients without diabetic retinopathy.

Wound healing activity of Salvia huberi ethanolic extract in streptozocin-induced diabetic rats.

Objective: The aim of this study was to examine the in vivo wound healing potential of Salvia huberi Hedge (endemic to Turkey) on excision and incision wound models in diabetic rats. Method: Male Wistar albino rats, 3-4 months old and weighing 180-240g were used. The animals were randomly divided into five groups including Control, Vehicle and Fito reference, and two different concentrations (0.5% and 1% weight/weight (w/w)) of ethanol extract of Salvia huberi were investigated in both wound models on streptozocin-induced diabetic rats using macroscopic, biomechanical, biochemical, histopathological, genotoxic and gene expression methods over both seven and 14 days. Fito cream (Tripharma Drug Industry and Trade Inc., Turkey) was used as the reference drug. Results: A total of 60 rats were used in this study. Salvia huberi ointments at 0.5% and 1% (w/w) concentrations and Fito cream showed 99.3%, 99.4% and 99.1% contraction for excision wounds, and 99.9%, 97.0% and 99% contraction for incision wounds, respectively. In Salvia huberi ointments and Fito cream groups, re-epithelialisation increased dramatically by both day 7 and day 14 (p<0.05). By day 14, low hydroxyproline and malondialdehyde (MDA) levels, and high glutathione (GSH) levels were observed in the Salvia huberi ointment groups. After two application periods, damaged cell percent and genetic damage index values and micronucleus frequency of Salvia huberi ointment treatment groups were lower than Control and Vehicle groups (p<0.001). A growth factor expression reached a high level by day 7 in the Control group; in Salvia huberi-treated groups it was decreased. Conclusion: The study showed that application of Salvia huberi ointments ameliorated the healing process in diabetic rats with excisional and incisional wounds and may serve as a potent healing agent.

Does asporin have a role in polycystic ovary syndrome? A pilot study.

OBJECTIVE: Asporin is secreted by theca cells in the mouse ovaries and is an effective marker at the gonadotropin-independent stage in secondary follicle development. It has an inhibitory effect on transforming growth factor beta and bone morphogenic proteins, which are involved in androgenesis process. Our aim was to compare serum asporin levels of polycystic ovary syndrome and control groups and examine the relationship between asporin and hyperandrogenism. METHODS: A total of 60 patients, i.e., 30 polycystic ovary syndrome group and 30 controls, were included in the study. The demographic characteristics, hormonal status, and serum asporin levels of patients were evaluated and compared for each group. In addition, polycystic ovary syndrome patients were analyzed according to the presence of hyperandrogenism. Receiver operating characteristic curve analysis was performed for asporin levels in order to distinguish polycystic ovary syndrome patients from controls. RESULTS: Body mass index, serum asporin and androgen levels, free androgen index, and insulin resistance values were statistically significantly higher in polycystic ovary syndrome group. Serum asporin levels were statistically significantly higher in hyperandrogenic polycystic ovary syndrome patients compared to non-hyperandrogenic polycystic ovary syndrome women (p=0.010). Receiver operating characteristic curve analysis was done for serum asporin levels to distinguish between polycystic ovary syndrome patients and healthy controls (area under the curve=0.676, standard error: 0.070, 95%CI: 0.539-0.812, p=0.019, 63.3% sensitivity, and 70% specificity). CONCLUSION: The elevation of serum asporin levels in patients with polycystic ovary syndrome may be associated with the pathogenesis of this syndrome, or it may be the consequence of the disease. This relationship may be explained through the androgen mechanism.

Does asporin have a role in polycystic ovary syndrome? A pilot study

SUMMARY OBJECTIVE: Asporin is secreted by theca cells in the mouse ovaries and is an effective marker at the gonadotropin-independent stage in secondary follicle development. It has an inhibitory effect on transforming growth factor beta and bone morphogenic proteins, which are involved in androgenesis process. Our aim was to compare serum asporin levels of polycystic ovary syndrome and control groups and examine the relationship between asporin and hyperandrogenism. METHODS: A total of 60 patients, i.e., 30 polycystic ovary syndrome group and 30 controls, were included in the study. The demographic characteristics, hormonal status, and serum asporin levels of patients were evaluated and compared for each group. In addition, polycystic ovary syndrome patients were analyzed according to the presence of hyperandrogenism. Receiver operating characteristic curve analysis was performed for asporin levels in order to distinguish polycystic ovary syndrome patients from controls. RESULTS: Body mass index, serum asporin and androgen levels, free androgen index, and insulin resistance values were statistically significantly higher in polycystic ovary syndrome group. Serum asporin levels were statistically significantly higher in hyperandrogenic polycystic ovary syndrome patients compared to non-hyperandrogenic polycystic ovary syndrome women (p=0.010). Receiver operating characteristic curve analysis was done for serum asporin levels to distinguish between polycystic ovary syndrome patients and healthy controls (area under the curve=0.676, standard error: 0.070, 95%CI: 0.539-0.812, p=0.019, 63.3% sensitivity, and 70% specificity). CONCLUSION: The elevation of serum asporin levels in patients with polycystic ovary syndrome may be associated with the pathogenesis of this syndrome, or it may be the consequence of the disease. This relationship may be explained through the androgen mechanism.

Biochemical, Histopathologic, and Genotoxic Effects of Ethanol Extract of Salvia hypargeia (Fisch. & Mey.) on Incisional and Excisional Wounded Diabetic Rats.

Purpose: Nonhealing wounds are a serious problem of diabetic patients. Salvia species are traditionally used for the treatment of wounds. The aim of the study was to investigate the effects of ointment prepared with ethanol extract obtained from the aerial parts of Salvia hypargeia, an endemic plant from Turkey, on diabetic rat incisional and excisional skin wounds. Materials and Methods: Male Wistar albino rats (n: 60) were divided into five groups. Diabetes was induced and two concentrations (0.5% and 1%) of the extract were used for ointments and applied on wounds for 7 and 14 days. Fito cream was chosen as a reference drug. Results: In excisional wounds, healing ratios of 0.5% (63.4% and 99.3%) and 1% (65.5% and 99.9%) S. hypargeia groups were higher compared to control (35.9% and 75.1%), and in incisional wounds, healing ratios of 0.5% (78.1% and 98.5%) and 1% (84.4% and 99.4%) S. hypargeia groups were higher compared to control (30.5% and 72.9%) (p < .01). Hydroxyproline (0.31 ± 0.3 and 0.34 ± 0.2) levels were lower and GSH (10.7 ± 3.1 and 7.6 ± 0.9) levels were higher in 0.5% and 1% S. hypargeia groups on the 14th day (p < .01). Histopathological results revealed re-epithelialization and formation of granulation tissue in all S. hypargeia groups. Genotoxicologic results indicated, GDI, DCP values, and MN frequency of 0.5% and 1% S. hypargeia groups did not reach to significant levels both on the 7 and 14 days. Conclusions: S. hypargeia may have a potential for therapeutic use in treatment and management of diabetic wounds with a successful topical application.

The effect of diabetes on vitreous levels of adiponectin and inflammatory cytokines in experimental rat model.

BACKGROUND: Diabetic retinopathy is one of the most common eye diseases faced by diabetic patients. It is a slow-progressing complication that results from damage to the blood vessels of the retina. OBJECTIVES: To investigate the role of adiponectin and inflammatory cytokines in the vitreous of diabetic rats. MATERIAL AND METHODS: The study was conducted in 3-4-month-old male albino Wistar rats (180-240 g). The animals were divided into 2 groups (n = 40 in each group): the diabetes group and the control group. A single dose of streptozotocin (STZ) (45 mg/kg) in citrate buffer (0.1 M; pH 4.5) was intraperitoneally (ip.) injected into the diabetes group rats. A single dose of citrate buffer was injected ip. into the control group rats. All subjects were sacrificed under intramuscular (im.) Na-thiopental (50 mg/kg) anesthesia. The rats' eyelids were opened with an eye speculum and vitreous samples were collected with 20G needles 4 mm posterior to the limbus. The levels of vitreous adiponectin, tumor necrosis factor α (TNF-α), interferon γ (INF-γ), and matrix metalloproteinase (MMP)-2 and -9 were determined using a solid-phase sandwich enzyme-linked immunosorbent assay (ELISA). RESULTS: The levels of adiponectin, TNF-α, INF-γ, MMP-2, and MMP-9 in the rat vitreous were significantly higher in the diabetes group than in the control group (p < 0.05). CONCLUSIONS: Elevated adiponectin, TNF-α, and INF-γ levels in the vitreous may be diagnostically useful in diabetic retinopathy, and inflammatory cytokines in the vitreous may be pathogenically important in this concentration.

Wound healing properties, antimicrobial and antioxidant activities of Salvia kronenburgii Rech. f. and Salvia euphratica Montbret, Aucher & Rech. f. var. euphratica on excision and incision wound models in diabetic rats.

Diabetic patients suffer from persistent and non-healing wounds. Salvia species are traditionally used for the treatment of wounds and colds. The aims of the present study were to evaluate the in vivo wound healing potential, in vitro antimicrobial and antioxidant activities, and total phenolic and flavonoid contents of the aerial parts of two endemic taxa, Salvia kronenburgii Rech. f. (SK) and Salvia euphratica Montbret, Aucher & Rech. f. var. euphratica (SE). Two different concentrations (0.5% and 1% (w/w)) of ethanol extracts were investigated in incision and excision wound models on Streptozotocin-induced diabetic rats using biomechanical, biochemical, histopathological, macroscopic, and genotoxic methods for 7 and 14 days. Antimicrobial activity was evaluated against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Acinetobacter baumannii, Aeromonas hydrophila, Mycobacterium tuberculosis, Candida glabrata, Candida parapsilosis, and Candida tropicalis using the broth microdilution and the resazurin microtiter assay plate methods. Fito®, Ampicillin, Ethambutol, Isoniazid, and Fluconazole were used as reference drugs. Antioxidant capacities and total phenolic and flavonoid contents of both extracts were detected using DPPH free radical scavenging assay, Folin-Ciocalteu, and Al(NO3)3 methods, respectively. SK ointment at 0.5% and 1% (w/w) concentrations and SE ointment at 1% (w/w) concentration showed 99.9%, 99.5%, and 99.7% contraction, respectively for excision wounds, and SK and SE ointments at 1% (w/w) concentration showed 99.4% and 99.2% contraction for incision wounds while Fito® showed 98.9% and 98.5% contraction, respectively. Increased re-epithelialization (P < 0.01 and P < 0.001), angiogenesis, and decreased dermal inflammation (P < 0.001) were determined for SK and SE ointments at both 7 and 14 days. SE ointment on day 7 and SK ointment on day 14 reduced oxidative damage to DNA when compared to control (P < 0.01 and P < 0.001). Both tested plants had greater antibacterial activity against A. baumannii (62.5 µg/mL MIC value) and SE had greater antimycobacterial activity against M. tuberculosis (0.24 µg/mL MIC value) when compared to reference drugs Ampicillin, Isoniazid, and Ethambutol (125, 0.97, and 1.95 µg/mL MIC values, respectively). Antioxidant capacities, total phenolic and flavonoid contents of SE and SK were 87.08%, 76.21 µg GAE/mg, 43.43 µg QE/mg and 72.17%, 41.81 µg GAE/mg, 33.62 µg QE/mg, respectively. SK and SE had strong wound healing effects while SK found to be more effective than SE at both 7 and 14 days.

Wound Healing Effect of Kaempferol in Diabetic and Nondiabetic Rats.

BACKGROUND: Flavonoids have previously been suggested to play a role in wound healing. To date, however, limited information is available on the wound healing effect of kaempferol (KM), which belongs to the class of flavonoids. The objective of this study was to determine the wound healing effects of KM. MATERIALS AND METHODS: The wound healing effects of KM with two different concentrations (0.5% and 1% [weight/weight, w/w]) were evaluated in incisional and excisional wound models on diabetic and nondiabetic rats by macroscopic, biomechanical, biochemical, and histopathological analyses. Diabetes was induced by streptozotocin. The KM ointments were prepared using a mixture of glycol stearate:propylene glycol:liquid paraffin (3:6:1); 0.5 g of the ointments were topically applied on the wounded areas once a day for 7 and 14 d. On days 0, 7, and 14, wounds were photographed, and macroscopic examination of the wounds was performed. After 7 and 14 d, hydroxyproline levels, biomechanical analysis, and histopathological parameters (reepithelialization, thickness of granulation tissue, angiogenesis, presence of inflammation, deposition of collagen, presence of fibrosis, degree of dermal inflammation, and number of mast cells) were assessed. RESULTS: The best wound healing effect was observed in the diabetic excisional and nondiabetic incisional wounds (92.12% and 94.17%, respectively) treated with 1% (w/w) KM ointment for 14 d according to macroscopic examination. The nondiabetic excisional (14th day) and incisional (7th day) wounds treated with 1% (w/w) KM ointment showed statistically higher levels of hydroxyproline than the control groups (2.84 and 2.07 µg/mg, respectively, P < 0.01). Reepithelialization scores of KM-treated diabetic and nondiabetic excisional wounds on both 7 and 14 d (P < 0.05 and P < 0.01) and incisional wounds on the day 14 (P < 0.05) were significantly higher than controls. The maximum tensile strength was observed in nondiabetic and diabetic groups (0.92 and 0.82 g/s, respectively) treated with 0.5% (w/w) KM ointment on day 14. CONCLUSIONS: Thus, KM appears to be an effective topical wound healing agent in the treatment of both nondiabetic and diabetic wounds.

Ultrastructural Alterations of Liver Tissue Cells in Methotrexate-Treated Balb/c Mice.

AIM: The current study investigated the efficacy of methotrexate (MTX) on liver tissue cells of Balb/c mice at the ultrastructural level using transmission electron microscopy. BACKGROUND: This agent is well known and used as a chemotherapeutic agent for a long time and not selective for cancer cells so, healthy cells beside cancer cells are also affected by MTX. MATERIALS AND METHODS: Experimental animals were divided into two groups; the first group was kept without treatment and served as the control, the second group was treated with 115 mg/kg MTX i.p. once weekly for 4 weeks and sacrificed under anesthesia after the 4th week. The liver tissues were osmium fixed and embedded in araldite, sectioned and observed under transmission electron microscope. RESULTS: Normal cell ultrastructure was determined in the control group whereas the liver cells of the MTX-treated group revealed ultrastructural alterations, such as the increase in lipid droplets, discontinuity of rough endoplasmic reticulum cisternae and vacuole formation. In addition, the loss of cytoplasmic material in hepatocytes was also evident. Condensation of nuclear chromatin and fusion of nucleic membranes were observed in the liver cells of the treated group. CONCLUSIONS: Results of the study indicated that MTX, used for different types of medical treatment, disturbed liver cell ultrastructure.

The prevalence of exfoliation syndrome in Turkey.

PURPOSE: To investigate the prevalence of the exfoliation syndrome and its relationship with ocular and cardiovascular diseases in the Central Anatolia region of Turkey. METHODS: This cross-sectional and population-based study was conducted at the Sivas Province among the population aged 40 years and over. The diagnosis of XFS was made when exfoliative material was found on the anterior lens capsule or iris on slit-lamp examination. The subjects were divided into an XFS group and a non-XFS group according to the presence of exfoliative material, and the groups were compared for the presence of glaucoma, cataract, age-related macular degeneration, phacodonesis, hypertension, diabetes mellitus, coronary artery disease, smoking and alcohol-use frequency. RESULTS: XFS was present in 63 subjects consisting of 42 males (8.0%) and 21 females (3.6%) for an overall rate of 5.7% (95% CI: 0.054-0.060). Once we adjusted the values for age, we found a statistically significant relationship of increased age and male gender with the presence of XFS (p = 0.001, p = 0.027, respectively). The relationship between XFS and glaucoma, cataract and phacodonesis was found to be statistically significant (p = 0.001). No relationship was found between exfoliation syndrome and hypertension, diabetes mellitus and coronary artery disease. CONCLUSION: The prevalence of exfoliation syndrome was 5.7% in this population-based study. There was a statistically significant relationship between XFS and advancing age and male gender.

Effects of Equisetum arvense Ointment on Diabetic Wound Healing in Rats.

UNLABELLED: This study is designed to evaluate the effect of Equisetum arvense (EA) ointment on dermal diabetic wound healing in streptozotocin-induced diabetic rats. METHODS: In this study, diabetes was induced via a single intraperitoneal injection of 65 mg/kg streptozotocin in 56 male Wistar rats. After anesthetization, a 15 mm x 15 mm wound for each rat was made by removing skin in a circle on the dorsum. A total of 56 diabetic wounds were studied in 8 groups (n = 7), 4 of which were treated with EA ointment. On the 7th and 14th days after creating the wounds, the state of the diabetic wound healing was evaluated with wound closure ratio and by performing histopathologic studies. RESULTS: Groups treated with EA 5%-10% ointment were found to have a statistically higher wound closure ratio than control and petroleum jelly-lanolin groups (P < 0.05). On day 14, groups to which EA 5%-10% ointment was applied showed 99.71% and 99.93% wound closure ratio (P < 0.05) and higher dermal and epidermal regeneration, angiogenesis, and granulation tissue thickness after 14 days than the other groups (P < 0.05). CONCLUSION: These results indicate that EA ointments exhibit significant diabetic wound healing activity in excision wounds. Further clinical and experimental studies are needed to confirm these results.

The effects of long-term exposure of magnetic field via 900-MHz GSM radiation on some biochemical parameters and brain histology in rats.

The aim of this study is to determine the effects of magnetic field via cell phones on some blood parameters and neurons in the brain of rats. Animals have been classified into three groups: control, Magnetic Field (MF), and F2 groups. Throughout this study, cell phones were placed on the wall of the cages. Rats were exposed to the effects of cell phones during prenatal and postnatal periods until they were 80 days old. During the study, the exposure procedure of rats was that the phone was in standby mode for a whole day and in talking mode for 30 min per day. The waves of cell phones caused an increased blood glucose level from 96.52 ± 5.64 mg/dl to 132.14 ± 5.93 mg/dl and an increased serum protein level from 131.14 ± 6.19 mg/dl to 319.29 ± 6.73 mg/dl compared to control. Statistically, significant differences wasn't observed in the blood cholesterol concentration between the groups compared to the control. Weekly weight gain decreased in all groups compared to the control. MF exposure decreased pyramidal neuron numbers 51.15% and increased ischemic neuron numbers 73% at cortex region of brain. In addition, vascular dilatations have increased clearly in group F2.Whereas the procedure of MF did not have any effects on hippocampal pyramidal cell numbers, magnetic fields increased the amount of ischemic neurons three-fold compared to the control. In conclusion, MF affected some biochemical parameters, especially the cortex region of the brain.

Effects of Equisetum arvense Ointment on Dermal Wound Healing in Rats .

UNLABELLED: The aim of this study was to evaluate the effectiveness of Equisetum arvense ointment on dermal wound healing in rats. METHODS: The authors studied a total of 56 wounds in four groups of rats. Each wound measured 15 mm x 15 mm. The first group did not receive treatment while the second group was treated with a 1:1 mixture of Vaseline and lanolin ointment. Equisetum arvense ointment doses of 5% and 10% were used in the third and fourth groups. The rats were observed at days 7 and 14 post wounding. The state of the wound healing was evaluated using wound closure ratio and histopathologic studies. RESULTS: Equisetum arvense 5% and 10% groups and the Vaseline-lanolin group had a statistically significant higher wound closure ratio than the control group (P &0.05). Equisetum arvense ointment groups had a 95.26% and 99.96% wound closure ratio (P & 0.05) and higher dermal and epidermal regeneration, angiogenesis, and granulation tissue thickness after 14 days as compared to the other groups (P & 0.05). P. Equisetum arvense ointment exhibits significant wound healing activity in rats. Further clinical and experimental studies are needed to confirm these results. .

Topical atorvastatin in the treatment of diabetic wounds.

BACKGROUND: Currently, it is reported that statins may be useful in the treatment of diabetes mellitus foot ulceration. The aim of this study was to evaluate treatment of the wounds in streptozotocin-induced diabetic rats with local atorvastatin. METHODS: Two 15 x 15 mm-sized wounds were created in 28 streptozotocin-induced rats. A total of 56 diabetic wounds were studied in 8 groups (n = 7). No treatment was administered in the first and second groups, which lasted for 7 and 14 days, respectively. Third and fourth groups consisted of diabetic rats that were administered 1:1 mixture of lanolin and vaseline therapy for 7 and 14 days, respectively. One percent statin plus 1:1 mixture of lanolin and vaseline was used in the fifth and sixth groups for 7 and 14 days, respectively; and in seventh and eighth groups, 5% statin plus 1:1 mixture of lanolin and vaseline therapy was used for 7 and 14 days, respectively. On the 7th and 14th days, state of the wound healing was observed, and the percent of wound healing was determined by measuring its size and by performing a histopathologic study. The statistical analyses were performed by Mann-Whitney U test, using SPSS 14.0 software. RESULTS: On the 14th day, the rates of wound healing in the first, second, third, and fourth groups were 14%, 40%, 96.59%, and 96.51%, respectively. This ratio was calculated by the formula healing ratio (%) = 100 x (1-wound area/initial wound area). Accordingly, in the multiple comparisons, the rates of wound healing were found to be significantly higher in the diabetic rat groups administered 1% and 5% atorvastatin compared with those administered a mixture of lanolin-vaseline and the untreated group (for comparison each one P < 0.001). CONCLUSIONS: Local atorvastatin therapy may be useful for healing the wounds in diabetic rats. Further clinical and experimental studies are needed to confirm these results.

Does long-term talc exposure have a carcinogenic effect on the female genital system of rats? An experimental pilot study.

OBJECTIVE: In several studies, the prolonged exposure to talc has been associated with development of ovarian cancer. However, some studies have advocated contrary views. The present study aims to investigate histopathological changes and whether long-term talc exposure is associated with potential carcinogenic effects on the female genital organs of rats. MATERIALS AND METHODS: The present study was conducted at Dumlupinar University Medical Faculty and a total of 28 Sprague-Dawley rats were included. The experimental animals were allocated into four groups having seven rats each. Groups 1 and 2 served as controls, where the rats in Group 1 did not receive any intervention and Group 2 received intravaginal saline. Groups 3 and 4 received intravaginal or perineal talc application, respectively. Talc was applied for 3 months on a daily basis. Histopathological changes in the peritoneum and female genital system were evaluated. For statistical analyses, Fisher's exact test was carried out using SPSS. FINDINGS: In both the groups exposed to talc (Groups 3 and 4), evidence of foreign body reaction and infection, along with an increase in inflammatory cells, were found in all the genital tissues. Genital infection was observed in 12 rats in the study group and 2 rats in the control group. Neoplastic change was not found. However, there was an increase in the number of follicles in animals exposed to talc. No peritoneal change was observed. In the groups not exposed to talc, similar infectious findings were found, but there was a statistically significant difference between the groups (Groups 1 and 2 vs. Groups 3 and 4, P > 0.05). Neoplastic change was also not observed in these groups. Four groups were compared in terms of neoplastic effects and infections. In Groups 1, 5 rats were normal, two developed vulvovaginitis and endometritis with overinfection (in both ovaries), and one developed salpingitis (in both fallopian tubes), that is, infection was found in a total of two rats. In Group 2, only one experimental animal had endometritis. All the animals in Groups 3 and 4 developed infections. CONCLUSIONS: Talc has unfavorable effects on the female genital system. However, this effect is in the form of foreign body reaction and infection, rather than being neoplastic.