Prevention of ovarian hyperstimulation syndrome in a rat model: comparison of the efficacy of tocilizumab with that of ranibizumab, cabergoline, and a gonadotropin-releasing hormone antagonist.

The aim of the study is to investigate the effects of the interleukin-6 (IL-6) blocker tocilizumab in a hyperstimulated rat model and compare it with ranibizumab, a gonadotropin-releasing hormone antagonist (GnRHA), and cabergoline. Forty-seven rats were randomly divided into the following seven groups: Group 1: OHS; Group 2: OHS+ GnRHA; Group 3: OHS + ranibizumab; Group 4: OHS + cabergoline; Group 5: OHS + low-dose tocilizumab (TL); Group 6: OHS + high-dose tocilizumab (TH); Group 7: sham. Ovarian weight was significantly lower only in the ranibizumab group than in the OHS group. Estrogen levels were significantly lower in the GnRHA group than in the OHS and the treatment groups. Progesterone levels were significantly lower in the ranibizumab, cabergoline, and TL groups than in the OHS group. Among the treatment groups, corpus luteum counts were lower than in the OHS group. Corpus luteum counts were lowest in the tocilizumab groups. IL-6 intensity was lower in all treatment groups than in the OHS group. In the ranibizumab group IL-6 intensity was the lowest. The TL group did not significantly differ from the GnRHA and cabergoline groups regarding IL-6 expression. Ovarian VEGF expression was significantly lower in all treatment groups. For the TL, ranibizumab, and cabergoline groups VEGF intensity was similar. Tocilizumab may be a new strategy for preventing ovarian hyperstimulation syndrome by inhibition of IL-6.

Single dose varenicline may trigger epileptic activity.

Varenicline is a new drug for smoking cessation, and its effect on epilepsy is not clear. The aim of this study was to investigate whether different doses of varenicline cause epileptic activity. Forty rats were randomly assigned to the following eight groups: control, saline, and 0.025, 0.04, 0.1, 0.5, 1, and 2 mg kg(-1) varenicline (single dose, i.p.). EEGs were recorded before the varenicline injection and during the following 240 min. While epileptic discharges were observed on the EEGs of the rats in all of the varenicline-treated groups, motor findings of epileptic seizure were not observed in some rats in these groups except the 1 and 2 mg kg(-1) groups. These findings indicate that different single doses of varenicline cause epileptic activity in rats.

Blood pressure measurement in freely moving rats by the tail cuff method.

Inconsistency in consecutive blood pressure values is one of the most frequently observed problems in tail cuff method. The aim of this study was to measure blood pressure using the tail cuff method in rats without heating, anesthesia, and movement restriction. In this study, it has been shown that blood pressure measurement could be obtained without problem using the tail cuff method in freely moving rats in their cage environment. Also, the reliability of consecutive blood pressure values obtained from freely moving rats was higher than ether anesthesia and restricted groups.

The effect of L-carnosine on erythrocyte deformability and aggregation according to the cell age in young and aged rats.

This study aimed to investigate alterations in hemorheology induced by L-carnosine, an anti- oxidant dipeptide, and to determine their relationship to oxidative stress in density-separated erythrocytes of aged and young rats. 28 male Sprague Dawley rats were divided into 4 groups as aged (Aca), young (Yca) L-carnosine groups (250 mg/kg L-carnosine, i.p.) and aged (As), young (Ys) control groups (saline, i.p.). Density separation was further performed to these groups in order to separate erythrocytes according to their age. Blood samples were used for the determination of erythrocyte deformability, aggregation; and oxidative stress parameters. Erythrocyte deformability of Yca group measured at 0.53 Pa was lower than Aca group. Similarly, deformability of least-dense (young) erythrocytes of Yca group was decreased compared to least-dense erythrocytes of Aca groups. Total antioxidant capacity (TAC) of Aca group was higher and oxidative stress index (OSI) lower than As group. Although L-carnosine resulted in an enhancement in TAC of aged rats, this favorable effect was not observed in erythrocyte deformability and aggregation in the dose applied in this study.

Hemorheological responses to progressive resistance exercise training in healthy young males.

BACKGROUND: This study aimed to explore the effects of progressive resistance exercise training (PRET) on hemorheology. MATERIAL/METHODS: Exercise sessions included 1-3 sets of 8-12 repetitions at 40-60% of 1-repetition maximum (1-RM) for 3 weeks and at 75-80% of 1-RM during weeks 4-12. Red blood cell (RBC) deformability and aggregation were determined by ektacytometry, plasma and whole blood viscosities (WBV) by rotational viscometry. Lactate concentration was evaluated by an analyzer and fibrinogen was evaluated by coagulometry. Plasma total oxidant/antioxidant status was measured by colorimetry. RESULTS: Following an acute increase after exercise on the first day, RBC deformability was elevated during weeks 3 and 4 (p=0.028; p=0.034, respectively). The last exercise protocol applied in week 12 again caused an acute increase in this parameter (p=0.034). RBC aggregation was increased acutely on the first day, but decreased after that throughout the protocol (p<0.05). At weeks 4 and 12 pre-exercise measurements of WBV at standard hematocrit and plasma viscosity were decreased (p=0.05; p=0.041, respectively), while post-exercise values were increased (p=0.005; p=0.04, respectively). Post-exercise WBV at autologous hematocrit measured at week 12 was increased (p=0.01). Lactate was elevated after each exercise session (p<0.05). Fibrinogen was decreased on the third week (p<0.01), while it was increased on the 4th week (p=0.005). Plasma antioxidant status was increased at week 3 (p=0.034) and oxidative stress index was decreased at week 4 (p=0.013) after exercise. CONCLUSIONS: The results of this study indicate that PRET may have positive effects on hemorheological parameters.

Surgical Procedures Required for Measurement Reduce Nerve Conduction Velocity: An In Vivo and In Vitro Comparative Study.

Background: Although in vitro methods have disadvantages, they are still commonly used to measure nerve conduction velocity (NCV) in experimental studies. Therefore, this study was designed to demonstrate the effect of the surgical procedures required for in vitro methods on nerve fibers and the effect of in vivo and in vitro methods on the results of electrophysiological measurements.Methods: Rats were assigned to the in vivo (control-1, injury-1, and diabetic-1) and in vitro (control-2, injury-2, and diabetic-2) groups. The NCV and compound action potential amplitudes were measured, and the nerve fibers were histologically examined.Results: Damaged axons and myelin sheaths were observed in the control-2 group. The electrophysiological values of the in vitro groups were lower than those of the in vivo groups. Furthermore, these values were lower for the diabetic and injury groups than for the control groups.Conclusions: This study showed that the surgical procedures required for the in vitro method reduced the measured values. Owing to the previous and current disadvantages of the in vitro method, the in vivo method was more sensitive for the NCV measurement. Moreover, measurements can be performed using the current in vivo method for small nerve fibers.

The effects of in vivo and ex vivo various degrees of cold exposure on erythrocyte deformability and aggregation.

BACKGROUND: This study aimed to investigate alterations in hemorheology by cold exposure, in vivo and ex vivo, and to determine their relationship to oxidative stress. MATERIAL/METHODS: Rats were divided into 2 in vivo and ex vivo cold exposure groups. The in vivo group was further divided into control (AR), AC (4°C, 2 hours) and ALTC (4°C, 6 hours) subgroups; and the ex vivo group was divided into control (BR) and BC (4°C, 2 hours) subgroups. Blood samples were used for the determination of erythrocyte deformability, aggregation, and oxidative stress parameters. RESULTS: Erythrocyte deformability and aggregation were not affected by 2-hour ex vivo cold exposure. While 2 hour in vivo cold exposure reduced erythrocyte deformability, it returned to normal after 6 hours, possibly due the compensation by acute neuroendocrine response. Six hours of cold exposure decreased aggregation index, and might be an adaptive mechanism allowing the continuation of circulation. Aggregation of ex vivo groups was lower compared to in vivo groups. Cold exposure at various temperatures did not cause alterations in plasma total oxidant antioxidant status and oxidative stress index (TOS, TAS, OSI) when considered together. CONCLUSIONS: Results of this study indicate that the alterations observed in hemorheological parameters due to cold exposure are far from being explained by the oxidative stress parameters determined herein.

The effects of docosahexaenoic acid supplementation and exercise on growth hormone and insulin-like growth factor I serum levels during chronic hypoxia in rats.

BACKGROUND: In this study we examined the effects of docosahexaenoic acid (DHA) on growth hormone (GH), insulin-like growth factor I (IGF-I) and insulin-like growth factor binding protein-3 (IGFBP-3) in response to chronic hypoxia and exercise training in hypoxic conditions. METHODS: Thirty-five rats were divided into five groups; control group (C), hypoxia group (H), hypoxia-exercise group (HE), hypoxia-docosahexaenoic acid group (HD), hypoxia-exercise-docosahexaenoic acid group (HED). A treadmill exercise was performed as 30 m/min for 20 min/day, 5 days per week for 28 days at level grade for the exercising groups (HE and HED). DHA was given to the HD and HED groups every day orally (36 mg/kg). The animals, except for the C group, were exposed to hypoxia for 28 days. RESULTS: Serum levels of GH and IGF-I in the H group decreased after chronic hypoxia (p<0.001). GH and IGF-I in the HD group also decreased compared with the C group (p<0.05, p<0.01, respectively). GH in C group did not show significant difference compared with the HE and HED groups. Decreased serum level of IGF-I was observed for the HED group (p<0.05). CONCLUSIONS: According to our findings, chronic hypoxia exposure decreases serum levels of GH, and IGF-I and exercise training have a slightly positive effect on GH/IGF-I axis during hypoxia. In addition, DHA supplementation slightly increases GH and IGF-I serum levels in hypoxic conditions. However, this effect on GH/IGF-I axis during hypoxia is not strong compared with exercise. Therefore, we concluded that exercise and/or DHA supplementation does not have additional positive effect on these hormones in hypoxic conditions.

Effect of sulfite treatment on erythrocyte deformability in young and aged rats.

It is known that aging is associated with marked effects on integrity and function of cell membrane. These effects may also be exacerbated by exogenous chemicals, e.g. sulfite. Thus, the aim of this paper is to examine the influence of sulfite on hemorheological and related hematological parameters in rats of various ages. In this study, male Wistar rats at the age of 3 and 18 months were used and the following parameters were evaluated: Mean Cell Volume (MCV), Mean Corpuscular Hemoglobin Concentration (MCHC), Red blood Cell (RBC) deformability and aggregation. The results show that aging is associated with a decrease in RBC deformability and MCHC, an increase in MCV. Sulfite administration significantly increased RBC deformability in both young and aged rats. Although MCHC was decreased in young rats, it was increased in aged rats in response to sulfite exposure. Additionally, sulfite induced a decrement in MCV of aged rats. Neither aging nor sulfite treatment caused significant alterations in RBC aggregation parameters in all experimental groups. In conclusion, these findings suggest that RBC deformability impairs with age and sulfite has ameliorating effects on RBC deformability in both young and aged rats.

Effect of thrombocytapheresis on blood rheology in healthy donors: role of nitric oxide.

Platelet transfusions are increasingly being used to treat thrombocytopenic conditions. Because of anticoagulation, changes in blood composition and extracorporeal circulation, donor apheresis may cause alterations in hemorheology. This study aimed at investigating the effects of thrombocytapheresis on donor blood rheology. The effect of nitric oxide (NO) on donor red blood cell (RBC) deformability after thrombocytapheresis was also studied. Platelets were collected by a Haemonetics MCS 3p cell seperator. Blood samples were obtained before and 15 min after thrombocytapheresis. RBC deformability and aggregation were measured using an ektacytometer, whole blood viscosity (WBV) was determined with a cone-plate rotational viscometer. Donor RBCs were shown to be less deformable at all stress levels except 0.30 Pa after thrombocytapheresis and NO donor sodium nitroprusside (SNP, 10(-6) M) reversed the reduced deformability caused by thrombocytapheresis. It was observed that donor apheresis induces a decrement in RBC aggregation and WBV measured at standard hematocrit (Hct). No significant alterations were observed in WBV values determined at native Hct values. Thrombocytapheresis also resulted in a decrement in fibrinogen, total protein, cholesterol and albumin levels whereas Hct was found to be increased and serum glucose, triglyceride, hemoglobin levels unaltered after apheresis. These results suggest that, thrombocytapheresis causes alterations in hemorheological parameters and hence in the perfusion of the microvasculature of the donors and NO appears to have a protective effect on the impairment observed in RBC deformability.

The effects of ex vivo ozone treatment on human erythrocyte carbonic anhydrase enzyme.

Ozone autohemotherapy is used in the treatment of some diseases. Carbonic anhydrases (CAs, EC 4.2.1.1) are metalloenzymes and play a role in homeostatic mechanisms. The aim of this study was to investigate the effects of ozone on human red blood cell CA (hCA) enzyme activity. Blood samples were treated with different doses of ozone (10, 20, 30 µg/ml) and the erythrocyte total CA activities were determined. Also, purified hCAI and hCAII isozymes were treated with the same doses of ozone and the enzyme activities were measured. About 30 µg/ml ozone treatment decreased the purified hCAI and hCAII activity and increased the total CA activity compared to the control. Because the implication of CAs on many physiological and biochemical processes is linked to pathologies, it can be suggested that the ozone at a concentration of 30 µg/ml is safely used by autohaemotherapy in a well-designed clinical trial.

The effects of air pollution and smoking on placental cadmium, zinc concentration and metallothionein expression.

This study is designed to determine the placental zinc (Zn) and cadmium (Cd) levels in mothers who were smokers, mothers who were thought to be exposed to air pollution, and mothers who were non-smokers and to investigate the relationship between the expression of placental metallothionein (MT) binding these metals and blood progesterone level. Placental Zn and Cd levels were measured by atomic absorption spectrometry. Presence of placental MT was determined immunohistochemically. Placental changes were examined by light microscope after H&E and PAS staining. Immunohistochemical MT staining of syncytiotrophoblastic and villous interstitial cells were scored as positive or negative. Among the 92 mothers included in the study, 33 were smokers (Group I), 29 had been exposed to air pollution (Group II) and 30 were non-smoker rural residents who had never been exposed to air pollution (Group III). Mean off-spring birth weight of 3198.62+/-380.01 g and mean placenta weight of 561.38+/-111.55 g of Group II were lower when compared with those of other two groups. In Group I, mean placental Cd and Zn were 0.063+/-0.022 microg/g and 39.84+/-15.5 microg/g, respectively, being higher than in other groups. In Group II, mean placental Cd and Zn levels were higher than those of Group III. Blood progesterone levels of subjects in Group I (121 ng/ml) were the lowest of all groups. While the mean count of villi was the highest in Group III; the highest mean count of syncytial knots was in Group II. Thickening of vasculo-syncytial membrane was most prominent in Group I. Similarly, MT staining was positive and very dense in 72.7% (24/33) of cases in Group I (p

Effect of penicillin G-induced epileptic seizures on hemorheological parameters in rats.

Normally, cerebral blood flow (CBF) is quantitatively coupled to cerebral metabolic rate like other tissues and maintained basically by altering vascular geometry and appropriate perfusion pressure. However, the rheological properties of the blood are important factors for effective tissue perfusion. Although a lot of studies have reported that hemorheological parameters are affected by a wide range of pathophysiological conditions, to our knowledge no research related to the effects of epileptic seizures on hemorheological parameters has been carried out. Thus, the aim of this study was to explore possible changes in rheological parameters including red blood cell (RBC) deformability, rigidity and aggregation, whole blood and plasma viscosity during epileptic seizures induced by penicillin G in rats. Eighteen female albino rats were divided into three groups that included sham operated controls (Group S), epileptic group (Group E), intraperitoneal penicillin group (Group IPP). Epilepsy was induced by intracortical injections of penicillin G. Hemorheological studies had been carried out 3 h after the induction of epilepsy. Among the studied hemorheological parameters, only RBC deformability was found to be different in the E group compared to S group. Epileptic seizures led to an increase in RBC deformability in the E group. In conclusion, these results suggest that in addition to an increase in CBF, RBC deformability may also improve to better match brain metabolic demands during seizures.

The effects of ozone therapy on caspase pathways, TNF-α, and HIF-1α in diabetic nephropathy.

BACKGROUND: Accelerated apoptosis plays a vital role in the development of diabetic vascular complications. Ozone may attenuate diabetic nephropathy by means of decreased apoptosis-related genes. The aim of our study was to investigate the effect of ozone therapy on streptozotocin-induced diabetic nephropathy in rats. Also the histopathological changes in diabetic kidney tissue with ozone treatment were evaluated. METHODS: The rats were randomly divided into six groups (n = 7): control (C), ozone (O), diabetic (D), ozone-treated diabetic (DO), insulin-treated diabetic (DI), and ozone- and insulin-treated diabetic (DOI). D, DI, and DOI groups were induced by a single intraperitoneal injection of streptozotocin. Ozone was given to the O, DO, and DOI groups. Group DI and DOI received subcutaneous (SC) insulin (3 IU). All animals received daily treatment for 6 weeks. RESULTS: Expressions of caspase-1-3-9, HIF-1α, and TNF-α genes were significantly higher in D group compared to C group (p < 0.05 for all). Ozone treatment resulted in significant decrease in the expressions of these genes in diabetic kidney tissue compared to both C and D group (p < 0.05 for all). Caspase-1-3-9, HIF-1α, and TNF-α gene expressions were found to be lower in DOI group compared to C group (p < 0.05 for all). Also adding ozone treatment to insulin therapy resulted in more significantly decrease in the expressions of these genes in diabetic tissue compared to only insulin-treated diabetic group (p < 0.05 for all). Regarding histological changes, ozone treatment resulted in decrease in the renal corpuscular inflammation and normal kidney morphology was observed. Both insulin and ozone therapies apparently improved kidney histological findings with less degenerated tubules and less inflammation of renal corpuscle compared to D, DO, and DI groups. CONCLUSION: Ozone therapy decreases the expressions of apoptotic genes in diabetic kidney tissue and improves the histopathological changes.

Evaluation of oxidative stress and erythrocyte properties in children with henoch-shoenlein purpura.

OBJECTIVE: Pathogenesis of Henoch-Schönlein purpura (HSP) is not clearly defined. The present study was conducted to investigate the alterations in erythrocyte deformability and oxidative stress in HSP and to examine the possible relationship between erythrocyte deformability and organ involvement in this disease. METHODS: Plasma malondialdehyde (MDA) levels, total antioxidant status (TAS), erythrocyte deformability and aggregation were measured in 21 children with HSP at the disease onset and during the remission period in comparison with healthy subjects. FINDINGS: HSP patients at the active stage had significantly higher MDA and lower TAS levels (P<0.05). Erythrocyte deformability was decreased at the active-stage and increased again at the remission period of HSP (P<0.05). Erythrocyte deformability was significantly decreased at four different shear stresses in patients with gastrointestinal system or renal involvement; and decreased at six different shear stresses in patients with gastrointestinal system, and renal involvement compared to the patients without organ involvement (P<0.05). No significant difference was observed in aggregation parameters (P>0.05). CONCLUSION: The present findings emphasize the association between impaired erythrocyte deformability and organ involvement in HSP.

The effects of anti-epileptic drugs on human erythrocyte carbonic anhydrase I and II isozymes.

Carbonic anhydrase (CA) is an enzyme which plays a role in various homeostatic mechanisms, such as acid-base balance and electrolyte secretion in a various tissues. This study was aimed at determining and comparing possible alterations in activity of this enzyme caused by the use of old (Carbamazepine, Phenytoin Sodium, Sodium Valproate) and new (Levetiracetam, Pregabalin, Gabapentin, Oxcarbazepine) anti-epileptic drugs. Blood samples were collected from the volunteers. The blood samples were centrifuged to separate plasma and erythrocyte package. Hemolysate was prepared from the red cells. CA I and II were purified from human erythrocytes by a simple one step procedure using Sepharose 4B-L-tyrosine-sulfonamide affinity column. CA I and II isozymes were treated with some anti-epileptic drugs, then the inhibition or activation of enzyme determined. The results of this study show that Levetiracetam is the most effective inhibitor for human erythrocytes carbonic anhydrase compared with the other anti-epileptic drugs.

Altitude training induced alterations in erythrocyte rheological properties: a controlled comparison study in rats.

Altitude training is frequently used by athletes to improve sea-level performance. However, the objective benefits of altitude training are controversial. This study aimed to investigate the possible alterations in hemorheological parameters in response to altitude training. Sprague Dawley rats, were divided into 6 groups: live low-train low (LLTL), live high-train high (LHTH), live high-train low (LHTL) and their controls live high and low (LHALC), live high (LHC), live low (LLC). LHC and LHTH groups were exposed to hypoxia (15% O2, altitudes of 3000 m), 4 weeks. LHALC and LHTL were exposed to 12 hours hypoxia/normoxia per day, 4 weeks. Hypoxia was maintained by a hypoxic tent. The training protocol corresponded to 60-70% of maximal exercise capacity. Rats of training groups ran on treadmill for 20-30 min/day, 4 days/week, 4 weeks. Erythrocyte deformability of LHC group was increased compared to LHALC and LLC. Deformability of LHTH group was higher than LHALC and LLTL groups. No statistically significant alteration in erythrocyte aggregation parameters was observed. There were no significant relationships between RBC deformability and exercise performance. The results of this study show that, living (LHC) and training at altitude (LHTH) seems more advantageous in hemorheological point of view.

Exercise and DHA prevent the negative effects of hypoxia on EEG and nerve conduction velocity.

It is known that hypoxia has a negative effect on nervous system functions, but exercise and DHA (docosahexaenoic acid) have positive effect. In this study, it was investigated whether exercise and/or DHA can prevent the effects of hypoxia on EEG and nerve conduction velocity (NCV). 35 adult Wistar albino male rats were divided into five groups (n=7): control (C), hypoxia (H), hypoxia and exercise (HE), hypoxia and DHA (HD), and hypoxia and exercise and DHA (HED) groups. During the 28-day hypoxia exposure, the HE and HED groups of rats were exercised (0% incline, 30 m/min speed, 20 min/day, 5 days a week). In addition, DHA (36 mg/kg/day) was given by oral gavage to rats in the HD and HED groups. While EEG records were taken before and after the experimental period, NCV records were taken after the experimental period from anesthetized rats. Data were analyzed by paired t-test, one-way ANOVA, and post hoc Tukey test. In this study, it was shown that exposure to hypoxia decreased theta activity and NCV, but exercise and DHA reduced the delta activity, while theta, alpha, beta activities, and NCV were increased. These results have shown that the effects of hypoxia exposure on EEG and NCV can be prevented by exercise and/or DHA.

Increased red blood cell deformability and decreased aggregation as potential adaptive mechanisms in the slow coronary flow phenomenon.

OBJECTIVE: At present, the precise pathophysiology of the slow coronary flow phenomenon (SCFP) is still unknown and there is no consensus as to how it should be treated. The rheological factors affect the clinical course of various cardiovascular diseases. We studied the intrinsic properties of blood in the SCFP. MATERIALS AND METHODS: Twenty-six SCFP patients who had angiographically confirmed SCFP, and had otherwise normal epicardial coronary arteries, were included in our study, as were 30 healthy individuals with normal results from arteriography. Red blood cell (RBC) deformability, aggregation, whole-blood viscosity at both native and standard (40%) hematocrit, and plasma viscosity were determined in each individual. The results were analyzed using a Mann-Whitney U-test, an unpaired t-test, and a χ-test, where appropriate. RESULTS: The mean thrombolysis in myocardial infarction frame count was significantly higher in SCFP patients than in the controls. RBC deformability measured at five different shear rates was significantly higher in SCFP patients than in the controls. The RBC aggregation index was lower in SCFP patients. There were no statistically significant differences in RBC aggregation half-time (t1/2) and aggregation amplitude, whole-blood viscosity, and plasma viscosity between the two groups. CONCLUSION: The SCFP is associated with increased RBC deformability and decreased RBC aggregation. These hemorheological alterations, possibly also contributing factors in limiting the pathogenesis, can especially serve as beneficial adaptive mechanisms in the SCFP.

Orexins increase penicillin-induced epileptic activity.

Orexins have been implicated with physiological function including sleep-wake cycle, energy homeostasis, drinking behavior, analgesia, attention, learning and memory but their effects on excitability are controversial. We investigated the effects of intracortical injections of orexin A (100 pmol) and B (100 pmol) on the electrophysiological manifestation of epileptic seizures induced by cortical penicillin application in adult male rats. In comparison to saline, orexin A and B enhanced significantly the spike number, spike amplitude and spectral power values induced by cortical penicillin. Our findings indicates that orexins enhances the hyperexcitable and hypersyncronic cortical epileptic activity induced by focal application of penicillin-G.