Ebru Balli

The Effect of Testosterone Replacement Therapy on Bladder Functions and Histology in Orchiectomized Mature Male Rats

OBJECTIVE To investigate the effect of testosterone replacement therapy on bladder functions and smooth muscle/collagen content in orchidectomized orchiectomized mature male rats. METHODS The study included 25 mature male Sprague-Dawley rats divided into 3 groups. After bilateral orchiectomy, 8 rats received intramuscular saline injection, as a control group, and 8 rats received intramuscular injection of testosterone undecanoate 100 mg/kg as a treatment group. The sham group had 9 rats. Urodynamic studies were performed in all groups, before and after the study. The rats were killed after 60 days, and cystometric findings and smooth muscle/collagen ratio of the bladders were compared between the groups. RESULTS From the beginning to the end of the experiment, mean maximal bladder capacity increased 46.61% +/- 20.82 in the testosterone treatment group, while decreased 38.91% +/- 17.83 in control group, revealing a significant difference (P = .002). Smooth muscle/collagen ratio was significantly higher in the testosterone treatment group (1.53 +/- .34) than in the control group (1.05 +/- .32), (P = .01). CONCLUSIONS This study showed that bladder capacity and smooth muscle/collagen content improved with testosterone therapy in orchiectomized rats. Therefore, testosterone replacement therapy in late-onset hypogonadal men with urogenital dysfunction may have a positive role to improve bladder function by increasing bladder smooth muscle.

The effect of testosterone alone and testosterone + estradiol therapy on bladder functions and smooth muscle/collagen content in surgically menopause induced rats

OBJECTIVES The aim of the study was to investigate the effect of testosterone alone and testosterone+estradiol therapy on bladder functions and smooth muscle/collagen content in surgically menopause induced rat model. METHODS The study included 34 female Sprague-Dawley rats, and the rats were divided into four groups. After bilateral oophorectomy, during a 60 days period, six rats received IM saline injection for one time, as a control group, and nine rats received testosterone undecanoate 100mg/kg IM for one time, and nine rats received testosterone undecanoate 100mg/kg IM for one time + daily 0.50mg nasal spray of 17beta estradiol. Ten rats were taken as sham group. Urodynamic studies were performed in all groups before and after the study. The rats were sacrificed after 60 days, and cystometric findings and smooth muscle/collagen ratio of the bladders were compared between the groups. RESULTS Increase in maximal bladder capacity and compliance were significantly higher in the testosterone treatment group and testosterone + estradiol treatment group than in the control group (p = 0.01 and p = 0.002, respectively for bladder capacity; p = 0.04 and p = 0.005, respectively for bladder compliance). Smooth muscle/collagen ratio of the bladders was significantly higher in the testosterone and testosterone + estradiol treatment groups than in the control group (p = 0.04 and p = 0.008, respectively). CONCLUSIONS This study shows that bladder functions may deteriorate in postmenopausal period. In addition to estrogen replacement therapy, testosterone has a significant role to increase bladder smooth muscle, leading to improvement in bladder functions in postmenopausal women with urogenital system dysfunction.

Exposure to gamma rays induces early alterations in skin in rodents: mechanical, biochemical and structural responses.

In this study, the effect of gamma rays has been investigated on the normal rat skin using biomechanical, biochemical and histological techniques. Seventeen male Wistar albino rats were divided into two groups (control (n=7) and irradiated (n=10)). The irradiated group was treated with a (60)Co gamma source at a dose of 10Gy at room temperature. Skin biomechanics were measured with tensile test using biomaterial testing machine and maximum load, stiffness, energy absorption capacity, ultimate stress, ultimate strain and elastic modulus were calculated. In the irradiated group, energy, strain and toughness were significantly lower than in the control group (p<0.05). However, strength, displacement, stiffness, stress and elastic modulus were similar to that of the control group (p>0.05). Catalase (CAT) activities and the levels of malondialdehyde (MDA) in the skin of rats were measured using the biochemical methods. MDA levels significantly increased whereas CAT activities decreased in the irradiated group as compared with the control group (p<0.05). Diameters of collagen fibers were measured by transmission electron microscopy. There was no significant difference (p>0.05) between control and irradiated groups for collagen fiber diameter. Thickness of epidermis was significantly lower than the control group. There were no changes in the epidermis between the irradiated group and the control group ultrastructurally. The results of this study show that the gamma irradiation has a significant effect on normal healthy skin.

Effects of levosimendan on myocardial ischaemia-reperfusion injury

Background and objective: Levosimendan has a cardioprotective action by inducing coronary vasodilatation and preconditioning by opening KATP channels. The aim of this study was to determine whether levosimendan enhances myocardial damage during hypothermic ischaemia and reperfusion in isolated rat hearts. Methods: Twenty‐one male Wistar rats were divided into three groups. After surgical preparation, coronary circulation was started by retrograde aortic perfusion using Krebs‐Henseleit buffer solution and lasted 15 min. After perfusion Group 1 (control; n = 7) received no further treatment. In Group 2 (non‐treated; n = 7), hearts were arrested with cold cardioplegic solution after perfusion and subjected to 60 min of hypothermic global ischaemia followed by 30 min reperfusion. In Group 3 (levosimendan treated; n = 7), levosimendan was added to the buffer solution during perfusion and the hearts were arrested with cold cardioplegic solution and subjected to 60 min of hypothermic global ischaemia followed by 30 min reperfusion. At the end of the reperfusion period, the hearts were prepared for biochemical assays and for histological analysis. Results: Tissue malondialdehyde levels were significantly lower in the levosimendan‐treated group than in the non‐treated group (P = 0.019). The tissue Na+‐K+ ATPase activity was significantly decreased in the non‐treated group than in the levosimendan‐treated group (P = 0.027). Tissue myeloperoxidase (MPO) enzyme activity was significantly higher in the non‐treated group than in the levosimendan‐treated group (P = 0.004). Electron microscopic examination of the hearts showed cardiomyocytic degeneration at the myofibril, mitochondria and sarcoplasmic reticulum in both non‐treated and levosimendan‐treated groups. The severity of these findings was more extensive in the non‐treated group. Conclusions: Treatment with levosimendan provided better cardioprotection with cold cardioplegic arrest followed by global hypothermic ischaemia in isolated rat hearts.

Determination of acute and chronic effects of cadmium on the cardiovascular system of rats

In this study, the systemic hemodynamics induced by acute and chronic cadmium (Cd+2) intoxication in the cardiovascular system of rats using thoracic electrical bioimpedance were examined and the acute and chronic effects of Cd+2 intoxication on the activities of antioxidant enzymes and malondialdehyde (MDA) were compared. Also, in this study, ultrastructural changes in the heart and aorta of rats were evaluated. Thirty-eight male Wistar albino rats were randomly divided into control, acute, and chronic groups. Chronic group was administered by oral gavage an aqueous solution of CdCl2 for 60 days, at dose of 15 mg Cd+2/kg/day. Acute group was administered by oral gavage an aqueous solution of CdCl2 with a single dose of 15 mg Cd+2/kg. Cadmium increased the stroke volume and cardiac output of rats in the chronic group, but did not change the heart rate significantly. Antioxidant enzymes activities and MDA level significantly increased in the chronic group. In ultrastructural examination, there were widespread degenerative changes in heart muscle cells of the chronic group but endothelial cells and smooth muscle cells in the aorta tissue samples had normal morphological features in all groups. All of the findings indicate that Cd+2 toxication can cause deformation in heart muscle cells due to an increase in free radicals and lipid peroxidation. Also, this study has confirmed that a long-term-Cd+2 exposure increased stroke volume (SV) and cardiac output (CO), but did not change the heart rate (HR).

Cyanoacrylate adhesive provides efficient local drug delivery.

Biodegradable drug delivery systems have advanced treatment of a wide spectrum of musculoskeletal problems. However, their lack of availability and cost can restrict use. To find an easily available and inexpensive biodegradable implant, we tested a widely used tissue adhesive, n-butyl-2-cyanoacrylate, as a drug-trapping material. We tested vancomycin with commercially available absorbable gelatin-sponge pieces as the scaffold. We evaluated the in vitro and in vivo drug release profiles and in vivo inflammatory response. A mouse muscle pouch model was used for in vivo evaluations. The released vancomycin level was measured by fluorescence polarization immunoassay technique, and a leukocyte count-based grading system was used to evaluate inflammatory response. Our findings suggest the proposed implant provides effective drug release for as much as 42 days in vitro and 14 days in vivo. The presence of n-butyl-2-cyanoacrylate led to a local inflammatory response which decreased after 3 weeks in the group with less adhesive. These results showed that n-butyl-2-cyanoacrylate could efficiently trap and slowly release a drug when used in the structure of a biodegradable local drug delivery device.

The Prevalence of Fabry Disease in Patients with Chronic Kidney Disease in Turkey: The TURKFAB Study.

Background/Aims: Fabry disease is a treatable cause of chronic kidney disease (CKD) characterized by a genetic deficiency of α-galactosidase A. European Renal Best Practice (ERBP) recommends screening for Fabry disease in CKD patients. However, this is based on expert opinion and there are no reports of the prevalence of Fabry disease in stage 1-5 CKD. Hence, we investigated the prevalence of Fabry disease in CKD patients not receiving renal replacement therapy. Methods: This prospective study assessed α-galactosidase activity in dried blood spots in 313 stage 1-5 CKD patients, 167 males, between ages of 18-70 years whose etiology of CKD was unknown and were not receiving renal replacement therapy. The diagnosis was confirmed by GLA gene mutation analysis. Results: Three (all males) of 313 CKD patients (0.95%) were diagnosed of Fabry disease, for a prevalence in males of 1.80%. Family screening identified 8 aditional Fabry patients with CKD. Of a total of 11 Fabry patients, 7 were male and started enzyme replacement therapy and 4 were female. The most frequent manifestations in male patients were fatigue (100%), tinnitus, vertigo, acroparesthesia, hypohidrosis, cornea verticillata and angiokeratoma (all 85%), heat intolerance (71%), and abdominal pain (57%). The most frequent manifestations in female patients were fatigue and cornea verticillata (50%), and tinnitus, vertigo and angiokeratoma (25%). Three patients had severe episodic abdominal pain attacks and proteinuria, and were misdiagnosed as familial Mediterranean fever. Conclusions: The prevalence of Fabry disease in selected CKD patients is in the range found among renal replacement therapy patients, but the disease is diagnosed at an earlier, treatable stage. These data support the ERBP recommendation to screen for Fabry disease in patients with CKD of unknown origin.

The effect of tumor necrosis factor-α inhibitor soon after hypoxia-ischemia on heart in neonatal rats

AIMS Perinatal hypoxic-ischemic insult has acute and long term deleterious effects on many organs including heart. Although tumor necrosis factor alpha (TNF-α) has been reported to increase soon after hypoxia, the inhibition of this mediator has not been documented. The aim of this study was to investigate the effects of a TNF-α inhibitor (etanercept) on contractility and ultrastructure of rat heart muscles exposed to hypoxia-ischemia during neonatal period. MAIN METHODS Forty-five seven-day old rats divided into three groups were included in this study. The right carotid arteries of Saline and Etanercept groups of rats were ligated and kept in a hypoxia chamber containing 8% oxygen for 2h. Immediately after hypoxia, while Etanercept group was administered 10mg/kg etanercept, Saline group had only saline intraperitoneally. The carotid arteries of rats in Sham group were located without ligation and hypoxia. Mechanical activity of heart was recorded and tissue samples were examined by electron microscopy in the sixteenth week following the hypoxia-ischemia. KEY FINDINGS While atrial contractile force in Etanercept group was similar to Sham group, there was significant decrease in Saline group (p<0.001). However, there was only non-significant decrease in ventricular contractility of Saline group comparing to Sham group (p>0.05). After hypoxia-ischemia, ultrastructural degenerative changes and mitochondrial damage in atriums of Etanercept group were significantly less severe than Saline group. SIGNIFICANCE This study demonstrated that neonatal hypoxia-ischemia caused long term cardiac dysfunction and ultrastructural degenerative changes in the heart of rats. TNF-α inhibitor administration soon after hypoxia-ischemia may have heart protective effect.

Optimal transport time and conditions for cartilage tissue samples and expanded chondrocyte suspensions.

For autologous chondrocyte implantation, the chondral tissue obtained is transferred from the operating room to the laboratory using specialized carrier systems within 24 hours. Similar expenses are used for the transport of cultured chondrocytes. The purpose of this study was to find the optimal temperature, size of tissue, and time that the chondrocytes can stand without losing viability and proliferative capacity. Fresh calf cartilage was harvested and divided into 24 groups. Half of the samples were diced into 1- to 2-mm(3) pieces. All 12 groups were kept at either 4 degrees C, 25 degrees C, or 37 degrees C for 1, 3, 5, or 7 days and were seeded for cell culture. Times to reach confluence values were compared. Produced cell suspensions were grouped similarly and tested similarly. Neither the temperature nor the waiting days caused any difference in the proliferative capacity of the cells. Diced tissues yielded a shorter time to reach confluence values. Chondral tissue obtained from the patient can be transferred to the laboratory at temperatures ranging from 4 degrees C to 37 degrees C in up to 7 days. These conditions did not affect the proliferative capacity or the viability of the chondrocytes. Dicing the tissue prior to transport will shorten total culturing time. The expanded cell suspensions should be transferred at temperatures from 4 degrees C to 25 degrees C within 3 days. Specialized carrier systems to get the chondral tissue from the operating room to the laboratory and to take the expanded chondrocytes back to the operating room within hours may not be necessary.

Clinical and histopathological relationship of sildenafil and bosentan treatments in rats with monocrotaline induced pulmonary hypertension

BACKGROUND Pulmonary arterial hypertension (PAH) is a challenging disorder characterized by increasing pulmonary artery pressure, which is hard to treat. OBJECTIVE This study was aimed to investigate the effects of bosentan, sildenafil and their combination. METHODS Saline or MCT were applied to Wistar rats. By the development of PAH (4th week), MCT-given rats were treated orally with bosentan, sildenafil and combination of sildenafil and bosentan or placebo. ECHO examinations were performed. Tissues obtained from all of the rats were evaluated under an electron microscope. RESULTS Left ventricular end diastolic diameter significantly increased in sildenafil and combined groups. Sildenafil group revealed a significant decrease in RV pressure and wall thickness. Examination of lung revealed a significant amount of connective tissue formation and increase in inflammatory cells in all the groups except controls in the interalveolar septum. Examination of PA revealed an increase in connective tissue volume, hypertrophic changes and expansions in granular endoplasmic reticulum cisternaes in smooth muscle cells in active groups rather than in the controls. Unlike the controls, the examination of the RV revealed an enlargement of the sarcoplasmic reticulum cisternaes in some cells, due to the calcium increase. CONCLUSION Sildenafil and the combined therapy demonstrated to have more impact on pressure and the RV parameters in rats, with lower inflammatory findings in lung tissue (Fig. 6, Ref. 31).