Disturbances In Rats Research Articles

Su1558 Stress Induced Different Psychological Disorders Showed Characteristic Patterns of Gut Microbiota Leading to Bowel Movement Disturbance in Rats

Su1558 Stress Induced Different Psychological Disorders Showed Characteristic Patterns of Gut Microbiota Leading to Bowel Movement Disturbance in Rats

Dynamics of cognitive disturbances in rats with acute cerebral ischemia on the background of introduction of 0.9 % solution NaCl

For today there are no clear recommendations on infusion therapy at the disease and injuries of brain, and infusion preparations are commonly used empirically. Within the framework of the complex study of an influence of the different infusion remedies on brain at an experimental ischemia of brain, an aim of this work is to investigate how the 0,9 % solution of NaCl influences on the dynamics of cognitive functions and neurological status of the rats with an acute cerebral ischemia at the course medical introduction into an animal organism. Methods . Experiments were carried out on 60 white rats-males. An acute disturbance of cerebral blood circulation (ADCBC) was modeled by means of the two-sided dressing of internal carotid arteries. The 0,9 % solution of NаСІ was injected intravenously in catheterized thigh vein 2,5 ml/kg 2 times/day (5 ml/kg for a day). The first introduction was carried out in 30 minutes after ADCBC and then every day in 12 hours during 7 days. The control groups consist of intact rats that received 0,9 % NаСІ and animals with a model ischemia without treatment. Neurological deficiency in animals was defined on the scale stroke-index McGrow C. P. The dynamics of position-finding activity was assessed in experiment “an open field”. An assessment of animal capacity to training and memorization of an aversive stimulus was studied in the test of conditioned response of passive avoidance. The results were processed using the program StatPlus 2009. Results . The study showed that bilateral carotid occlusion in rats without treatment is followed not only by the high animal lethality but also by the development of a hard neurological symptomatology and then by essential disturbance of mnestic functions in animals that survived during the recovery period of model insult (р<0,01). The study of lethality dynamics, neurological status, behavioral responses in rats with ADCBC on the model of bilateral carotid occlusion showed that the course therapeutic infusion of 0,9 % solution of NaCl had some advantages relative to the group of the control pathology on indicators of neurological deficiency, orientating and searching activity, capacity to training and memorization. Conclusions. Infusion therapy with 0,9 % solution of NaCl in rats with the model cerebral ischemia in some degree promotes the renewal of its cognitive functions (р<0,05) relative to the untreated animals. Therapeutic effect of infusion therapy obtained in experiment with isoosmolar 0,9 % solution of NaCl is a reason to study protective actions of infusion preparations of the other groups at an acute disturbance of cerebral blood circulation

Magnesium Supplementation Prevents and Reverses Experimentally Induced Movement Disturbances in Rats: Biochemical and Behavioral Parameters.

Reserpine administration results in a predictable animal model of orofacial dyskinesia (OD) that has been largely used to access movement disturbances related to extrapyramidal oxidative damage. Here, OD was acutely induced by reserpine (two doses of 0.7mg/kg subcutaneous (s.c.)), every other day for 3days), which was administered after (experiment 1) and before (experiment 2) magnesium (Mg) supplementation (40mg/kg/mL, peroral (p.o.)). In experiment 1, Mg was administered for 28days before reserpine treatment, while in experiment 2, it was initiated 24h after the last reserpine administration and was maintained for 10 consecutive days. Experiment 1 (prevention) showed that Mg supplementation was able to prevent reserpine-induced OD and catalepsy development. Mg was also able to prevent reactive species (RS) generation, thus preventing increase of protein carbonyl (PC) levels in both cortex and substantia nigra, but not in striatum. Experiment 2 (reversion) showed that Mg was able to decrease OD and catalepsy at all times assessed. In addition, Mg was able to decrease RS generation, with lower levels of PC in both cortex and striatum, but not in substantia nigra. These outcomes indicate that Mg is an important metal that should be present in the diet, since its intake is able to prevent and minimize the development of movement disorders closely related to oxidative damage in the extrapyramidal brain areas, such as OD.

Impact of β-adrenoceptor blockade on systemic inflammation and coagulation disturbances in rats with acute traumatic coagulopathy.

BackgroundSympathetic hyperactivity occurs early in acute traumatic coagulopathy (ATC) and is closely related to its development. β-adrenoceptor antagonists are known to alleviate adverse sympathetic effects and improve outcome in various diseases. We investigated whether β-blockers have protective effects against inflammation and endothelial and hemostatic disorders in ATC.Material/MethodsATC was induced in male Sprague-Dawley rats by trauma and hemorrhagic shock. Rats were randomly assigned to the sham, ATCC (ATC control), and ATCB (ATC with beta-adrenoceptor blockade) groups. Rats were injected intraperitoneally with propranolol or vehicle at baseline. Heart rate variability (HRV) and markers of inflammation, coagulation, and endothelial activation were measured, and Western blotting analysis of nuclear factor (NF)-κB was done after shock. Separate ATCC and ATCB groups were observed to compare overall mortality.ResultsHRV showed enhanced sympathetic tone in the ATCC group, which was reversed by propranolol. Propranolol attenuated the induction of pro-inflammatory cytokines TNF-α and IL-6, as well as fibrinolysis markers plasmin antiplasmin complex and tissue-type plasminogen activator. The increased serum syndecan-1 and soluble thrombomodulin were inhibited by propranolol, and the NF-κB expression was also decreased by propranolol pretreatment. But propranolol did not alter overall mortality in rats with ATC after shock.ConclusionsBeta-adrenoceptor blockade can alleviate sympathetic hyperactivity and exert anti-inflammatory, anti-fibrinolysis, and endothelial protective effects, confirming its pivotal role in the pathogenesis of ATC. Its mechanism in ATC should be explored further.

Effects of the 5-HT&lt;sub&gt;1A&lt;/sub&gt; Receptor Agonist Tandospirone on ACTH-Induced Sleep Disturbance in Rats

The aim of this study was to compare the effect of the serotonin (5-HT)1A receptor agonist tandospirone versus that of the benzodiazepine hypnotic flunitrazepam in a rat model of long-term adrenocorticotropic hormone (ACTH)-induced sleep disturbance. Rats implanted with electrodes for recording electroencephalogram and electromyogram were injected with ACTH once daily at a dose of 100 µg/rat. Administration of ACTH for 10 d caused a significant increase in sleep latency, decrease in non-rapid eye movement (non-REM) sleep time, and increase in wake time. Tandospirone caused a significant decrease in sleep latency and increase in non-REM sleep time in rats treated with ACTH. The effect of tandospirone on sleep patterns was antagonized by the 5-HT1A receptor antagonist WAY-100635. In contrast, flunitrazepam had no significant effect on sleep parameters in ACTH-treated rats. These results clearly indicate that long-term administration of ACTH causes sleep disturbance, and stimulating the 5-HT1A receptor by tandospirone may be efficacious for improving sleep in cases in which benzodiazepine hypnotics are ineffective.

Effect of Yokukansan on sleep disturbance in a rat model of cerebrovascular dementia

Aim: Symptoms of dementia are classified into cognitive dysfunction and behavioral and psychological symptoms of dementia, including sleep disturbance. Yokukansan (YKS) is effective for sleep disturbance in patients with dementia, but the mechanisms are still unclear. In this study, we evaluated sleep disturbance in rats with cerebral ischemia. We investigated the effect of YKS on sleep disturbance in cerebral ischemia-treated rats. Methods: Cerebral ischemia was induced by twice occluding both of the common carotid arteries. Wakefulness, non-rapid eye movement (NREM) sleep, and rapid eye movement (REM) sleep were classified using electroencephalography and electromyography. mRNA expression of sleep–wakefulness signals were quantified using reverse transcription–polymerase chain reaction. Results: Rats with cerebral ischemia had a higher total wakefulness time and lower total NREM sleep time during the light phase. These changes were ameliorated with 1000 mg/kg YKS for 14 days, but not 3 mg/kg donepezil for 7 days. mRNA expression of the prefrontal cortical prostaglandin (PG) E2 (EP4) receptor and of the PGD2 (DP) receptor was increased by cerebral ischemia. Elevated prefrontal cortical EP4 and DP receptor mRNA expression was reduced by YKS treatment. Conclusion: Rats with cerebral ischemia had NREM sleep disturbance. YKS might reduce this sleep disturbance by preventing neuroinflammation, and this was mediated by the EP4 and DP receptors in the prefrontal cortex.

Acid reflux directly causes sleep disturbances in rat with chronic esophagitis.

Background & AimsGastroesophageal reflux disease (GERD) is strongly associated with sleep disturbances. Proton pump inhibitor (PPI) therapy improves subjective but not objective sleep parameters in patients with GERD. This study aimed to investigate the association between GERD and sleep, and the effect of PPI on sleep by using a rat model of chronic acid reflux esophagitis.MethodsAcid reflux esophagitis was induced by ligating the transitional region between the forestomach and the glandular portion and then wrapping the duodenum near the pylorus. Rats underwent surgery for implantation of electrodes for electroencephalogram and electromyogram recordings, and they were transferred to a soundproof recording chamber. Polygraphic recordings were scored by using 10-s epochs for wake, rapid eye movement sleep, and non-rapid eye movement (NREM) sleep. To examine the role of acid reflux, rats were subcutaneously administered a PPI, omeprazole, at a dose of 20 mg/kg once daily.ResultsRats with reflux esophagitis presented with several erosions, ulcers, and mucosal thickening with basal hyperplasia and marked inflammatory infiltration. The reflux esophagitis group showed a 34.0% increase in wake (232.2±11.4 min and 173.3±7.4 min in the reflux esophagitis and control groups, respectively; p<0.01) accompanied by a reduction in NREM sleep during light period, an increase in sleep fragmentation, and more frequent stage transitions. The use of omeprazole significantly improved sleep disturbances caused by reflux esophagitis, and this effect was not observed when the PPI was withdrawn.ConclusionsAcid reflux directly causes sleep disturbances in rats with chronic esophagitis.

Doxorubicin-induced behavioral disturbances in rats: Protective effect of melatonin and captopril

ObjectiveDoxorubicin is a recognized chemotherapeutic agent widely employed in human malignancies. The limiting factor of its use is a number of side effects. The aim of this work was to show, whether administration of doxorubicin could induce behavioral disturbances in rats, and whether angiotensin-converting enzyme inhibitor captopril or melatonin can modify these potential alterations. Design and methodsFour groups of 3-month-old Wistar rats (twelve per group) were treated for 4weeks: control (placebo-treated), doxorubicin (DOX) (5mg/kg i.v. in a single intravenous dose), DOX rats treated with either melatonin (10mg/kg/24h) or captopril (100mg/kg/24h). Systolic blood pressure (SBP) and the level of oxidative stress were investigated and behavioral tests of anxiety—open field test (OF), elevated plus maze (EPM) and light–dark box (LDB) were accomplished. ResultsDoxorubicin increased significantly systolic blood pressure and parameters of oxidative stress. Moreover, doxorubicin enhanced the level of anxiety in the tests of OF, EPM, and LDB. Captopril and melatonin prevented the blood pressure rise and the enhancement of oxidative load. Importantly, both substances reduced the parameters of anxiety. ConclusionChronic administration of captopril or melatonin has shown anxiolytic effect in the model of doxorubicin‐induced anxiety. It does not seem unreasonable to suppose that this protective effect of captopril or melatonin against anxiety development might have been related to the antioxidative effects of both substances.

Effect of Zige lyophilized powder for injection in improving acute cerebral microcirculation disturbance in rats

To investigate the effect of Zige lyophilized powder for injection in improving the acute cerebral microcirculation disturbance in rats. Window craniotomy was performed for rats after the drug administration for 14 days. The experimental microcirculation disturbance model was duplicated with high molecule dextran. After the drug administration, the micro-vein diameters of cerebral pla mater of various groups were observed and recorded under the biological microscope. The blood flow volume was monitored by laser Doppler flow-meter. HCT was measured by the electric resistance method. The hemorheological indexes were detected by the auto-hemorheological instrument. Zige lyophilized powder for injection (16.40, 32.70, 65.40 mg x kg(-1)) could significantly expand the micro-vein diameter of cerebral pla mater, improve the downward trend of the blood flow volume, and reduce the various hemorheological indexes. Zige lyophilized powder for injection shows the effect in improving the cerebral microcirculation.

Maternal Obesity Caused by Overnutrition Exposure Leads to Reversal Learning Deficits and Striatal Disturbance in Rats

Maternal obesity caused by overnutrition during pregnancy increases susceptibility to metabolic risks in adulthood, such as obesity, insulin resistance, and type 2 diabetes; however, whether and how it affects the cognitive system associated with the brain remains elusive. Here, we report that pregnant obesity induced by exposure to excessive high fatty or highly palatable food specifically impaired reversal learning, a kind of adaptive behavior, while leaving serum metabolic metrics intact in the offspring of rats, suggesting a much earlier functional and structural defects possibly occurred in the central nervous system than in the metabolic system in the offspring born in unfavorable intrauterine nutritional environment. Mechanically, we found that above mentioned cognitive inflexibility might be associated with significant striatal disturbance including impaired dopamine homeostasis and disrupted leptin signaling in the adult offspring. These collective data add a novel perspective of understanding the adverse postnatal sequelae in central nervous system induced by developmental programming and the related molecular mechanism through which priming of risk for developmental disorders may occur during early life.

Neurodegeneration and inflammation in hippocampus in experimental autoimmune encephalomyelitis induced in rats by one – Time administration of encephalitogenic T cells

Cognitive dysfunction is relatively frequent in multiple sclerosis (MS) and it happens from the early stages of the disease. There is increasing evidence that the grey matter may be involved in autoimmune inflammation during relapses of MS. The purpose of this study was to evaluate if a single transfer of encephalitogenic T cells, mimicking a relapse of MS, may cause hippocampal damage and memory disturbances in rats. Lewis rats were injected with anti-MBP CD4+ T cells, that induced one-phase autoimmune encephalomyelitis (EAE) with full recovery from motor impairments at 10–15days. The spatial learning and memory were tested by the Morris water maze test in control and EAE animals, 30 and 90days post-induction (dpi). The neural injury and inflammation was investigated in the hippocampus by immunohistochemistry and quantitative analyses. There was a marked decrease in the number of CA1 and CA4 pyramidal neurons 5dpi. The loss of neurons then aggravated till the 90 dpi. An increase in microglial and astroglial activation and in pro-inflammatory cytokines mRNA expression in the hippocampus, were present 30 and 90dpi. Nerve growth factor and brain-derived neurotrophic factor mRNA levels were also significantly elevated. The water maze test, however, did not reveal memory deficits. The present data indicate that a single transfer of autoimmune T cells results in preserved inflammation and probable on-going neuronal injury in the hippocampus, long after recovery from motor disturbances. These findings suggest that any relapse of the MS may start the neurodegenerative process in the hippocampus, which is not necessarily connected with memory deficits.

Mortal hyperkalemia disturbances in rats are NO-system related. The life saving effect of pentadecapeptide BPC 157

We demonstrate the full counteracting ability of stable gastric pentadecapeptide BPC 157 against KCl-overdose (intraperitoneal (i), intragastric (ii), in vitro (iii)), NO-system related. (i) We demonstrated potential (/kg) of: BPC 157 (10ng, 10μg ip, complete counteraction), l-arginine (100mg ip, attenuation) vs. L-NAME (5mg ip, deadly aggravation), given alone and/or combined, before or after intraperitoneal KCl-solution application (9mEq/kg). Therapy was confronted with promptly unrelenting hyperkalemia (>12mmol/L), arrhythmias (and muscular weakness, hypertension, low pressure in lower esophageal and pyloric sphincter) with an ultimate and a regularly inevitable lethal outcome within 30min. Previously, we established BPC 157-NO-system interaction; now, a huge life-saving potential. Given 30min before KCl, all BPC 157 regimens regained sinus rhythm, had less prolongation of QRS, and had no asystolic pause. BPC 157 therapy, given 10min after KCl-application, starts the rescue within 5–10min, completely restoring normal sinus rhythm at 1h. Likewise, other hyperkalemia-disturbances (muscular weakness, hypertension, low sphincteric pressure) were also counteracted. Accordingly with NO-system relation, deadly aggravation by L-NAME: l-arginine brings the values to the control levels while BPC 157 always completely nullified lesions, markedly below those of controls. Combined with l-arginine, BPC 157 exhibited no additive effect. (ii) Intragastric KCl-solution application (27mEq/kg) — (hyperkalemia 7mmol/L): severe stomach mucosal lesions, sphincter failure and peaked T waves were fully counteracted by intragastric BPC 157 (10ng, 10μg) application, given 30min before or 10min after KCl. (iii). In HEK293 cells, hyperkalemic conditions (18.6mM potassium concentrations), BPC 157 directly affects potassium conductance, counteracting the effect on membrane potential and depolarizations caused by hyperkalemic conditions.

The activity of antiparkinsonian drug hemantane in models of peripheral inflammation and lipopolysaccharide-induced neuroinflammation

A large body of literature supports the idea that inflammation exacerbates neurodegenerative pathology. This idea is also supported by the fact that intracerebral or intraperitoneal injection of lipopolysaccharide (LPS) induces symptoms of Parkinson’s disease in rats. The aim of this study is to evaluate the anti-inflammatory effects of the novel antiparkinsonian drug hemantane (N-2(adamantyl)hexamethylenimine hydrochloride), which is currently undergoing clinical trials, in models of peripheral inflammation and neuroinflammation and to investigate its ulcerogenic action, which is a common side effect of nonselective nonsteroidal anti-inflammatory drugs. Acetic acid-induced peritonitis in mice was used as a model of peripheral inflammation. Effect on the stomach was investigated in rats were deprived of food for 16 hours and then were treated with 0.2 LD50 of hemantane or the comparator drug diclofenac sodium per os. Injection of LPS in the left substantia nigra pars compacta in rats was chosen as a model of neuroinflammation. LPS-induced body weight loss, forelimb akinesia and behavioral changes caused by irritating odor were registered in rats. Hemantane in the dosage range of 10 - 40 mg/kg demonstrates anti-inflammatory activity and significantly decreases the intensity of exudative reaction in a model of acetic acid-induced peritonitis in mice. Additionally, at the dose of 0.2 LD50 orally it did not damage the gastric mucosa of rats. In a model of neuroinflammation induced by a unilateral injection of LPS, hemantane (10 mg/kg) prevents weight loss, development of forepaw akinesia contralateral to the operation, and smell disturbance in rats. Effectiveness of hemantane in the animal models of peripheral inflammation and neuroinflammation make it possible to suggest a new application of hemantane as a safe anti-inflammatory drug.

Changes in glucose tolerance and leptin responsiveness of rats offered a choice of lard, sucrose, and chow

Rats offered chow, lard, and 30% sucrose solution (choice) rapidly become obese. We tested metabolic disturbances in rats offered choice, chow+lard, or chow+30% sucrose solution [chow+liquid sucrose (LS)] and compared them with rats fed a composite 60% kcal fat, 7% sucrose diet [high-fat diet (HFD)], or a 10% kcal fat, 35% sucrose diet [low-fat diet (LFD)]. Choice rats had the highest energy intake, but HFD rats gained the most weight. After 23 days carcass fat was the same for choice, HFD, chow+lard, and chow+LS groups. Glucose clearance was the same for all groups during an intraperitoneal glucose tolerance test (GTT) on day 12, but fasting insulin was increased in choice, LFD fed, and chow+LS rats. By contrast, only choice and chow+LS rats were resistant to an intraperitoneal injection of 2 mg leptin/kg on day 17. In experiment 2 choice rats were insulin insensitive during an intraperitoneal GTT, but this was corrected in an oral GTT due to GLP-1 release. UCP-1 protein was increased in brown fat and inguinal white fat in choice rats, and this was associated with a significant increase in energy expenditure of choice rats during the dark period whether expenditure was expressed on a per animal or a metabolic body size basis. The increase in expenditure obviously was not great enough to prevent development of obesity. Further studies are required to determine the mechanistic basis of the rapid onset of leptin resistance in choice rats and how consumption of sucrose solution drives this process.

High-fructose corn syrup causes vascular dysfunction associated with metabolic disturbance in rats: Protective effect of resveratrol

High-fructose corn syrup (HFCS) is used in many prepared foods and soft drinks. However, limited data is available on the consequences of HFCS consumption on metabolic and cardiovascular functions. This study was, therefore, designed to assess whether HFCS drinking influences the endothelial and vascular function in association with metabolic disturbances in rats. Additionally, resveratrol was tested at challenge with HFCS. We investigated the effects of HFCS (10% and 20%) and resveratrol (50mg/l) beverages on several metabolic parameters as well as endothelial relaxation, vascular contractions, expressions of endothelial nitric oxide synthase (eNOS), sirtuin 1 (SIRT1), gp91phox and p22phox proteins and superoxide generation in the aortas. Consumption of HFCS (20%) increased serum triglyceride, VLDL and insulin levels as well as blood pressure. Impaired relaxation to acetylcholine and intensified contractions to phenylephrine and angiotensin II were associated with decreased eNOS and SIRT1 whereas increased gp91phox and p22phox proteins, along with provoked superoxide production in the aortas from HFCS-treated rats. Resveratrol supplementation efficiently restored HFCS-induced deteriorations. Thus, intake of HFCS leads to vascular dysfunction by decreasing vasoprotective factors and provoking oxidative stress in association with metabolic disturbances. Resveratrol has a protective potential against the harmful consequences of HFCS consumption.

The Significance of Microthrombosis and fgl2 in No-Reflow Phenomenon of Rats With Acute Myocardial Ischemia/Reperfusion

No-reflow phenomenon due to cardiac microvascular dysfunction or disturbance aggravates clinic outcomes of a portion of patients with acute myocardial infarction undergoing percutaneous coronary intervention or thrombolytic therapy. Our working hypothesis was that cardiac microthrombosis would play an important role in the pathogenesis. We investigated that cardiac microthrombi were observed by Martius, Scarlet, Blue methocl (MSB) and Masson trichrome staining. Furthermore, we investigated the expression of fibrinogen-like protein 2 (fgl2) in rats with acute myocardial ischemia/reperfusion (MI/R) and its possible pathological and clinical significance. The fgl2 was highly expressed in myocardium of rats with acute MI/R and located at cardiac microvascular walls. We found that the expression of fgl2 in peripheral mononuclear cells of rats with acute MI/R significantly increased correspondingly with its cardiac expression. Expression of cardiac fgl2 was correlated with no-reflow size of rats with acute MI/R, which was detected and calculated by thioflavin S staining. No-reflow size was in line with cardiac diastolic dysfunction of rats with acute MI/R monitored by hemodynamics. Thus, microthrombosis is involved in cardiac microvascular dysfunction or disturbance of rats with acute MI/R as one cause, and fgl2 may emerge as a predictor of the occurrence of no-reflow phenomenon.

L-theanine partially counteracts caffeine-induced sleep disturbances in rats

L-theanine has been reported to inhibit the excitatory effects of caffeine. The present study examined the effects of L-theanine on caffeine-induced sleep disturbances in rats. Rats received the following drug pairings: saline and saline (Control), 7.5mg/kg caffeine and saline, or 7.5mg/kg of caffeine followed by various doses of L-theanine (22.5, 37.5, 75, or 150mg/kg). Vigilance states were divided into: wakefulness (W), transition to slow-wave sleep (tSWS), slow-wave sleep (SWS), and rapid-eye-movement sleep (REMS). Caffeine significantly increased the duration of W and decreased the duration of SWS and REMS compared to the Control. Although L-theanine failed to reverse the caffeine-induced W increase, at 22.5 and 37.5mg/kg (but not at 75 and 150mg/kg), it significantly reversed caffeine-induced decreases in SWS. In conclusion, low doses of L-theanine can partially reverse caffeine-induced reductions in SWS; however, effects of L-theanine on caffeine-induced insomnia do not appear to increase dose-dependently.

Alcohol‐induced Osteonecrosis – dose and duration effects

We read with great interest the paper by Ikemura et al. published in the August issue of the International Journal of Experimental Pathology (Ikemura et al. 2011). In their study, the authors used a model of alcohol-induced osteonecrosis in rabbits to evaluate the morphological changes in bone marrow fat cells and the changes in the serum lipid levels. Animals received either 15 ml/kg/day (LDA: low-dose alcohol) or 30 ml/kg/day (HDA: high-dose alcohol) of a solution containing 15% ethanol for intragastrically for 4 weeks, whereas a control group received a physiological saline solution. Results were observed at 6 weeks after the beginning of the treatment (i.e. 2 weeks after withdrawal). As these results are difficult to reconcile with aspects of current understanding of alcohol effects on bone metabolism, we would like to comment on their protocol and their results in perspective. Regarding the quantity of alcohol used to induce osteonecrosis, the animals thus ingested either 2.25 g/kg/day (LDA) or 4.5 g/kg/day (HDA) of alcohol. In comparison, Wang et al. (2008) used a dose of 9.2 g/kg/day (i.e. more than twice the amount used in the HDA group) to induce objectively recognizable alcohol-induced osteonecrosis in rabbits, with concomitant marked marrow fat cell hypertrophy and proliferation, thinner and sparse trabeculae, diminished hematopoiesis and increased empty osteocyte lacunae (Wang et al. 2008). In the same manner, Broulik et al.(2010) used a dose of 7.22 g/kg/day of ethanol (corresponding to a consumption of 1 l wine or 2.5 l of 12 ° beer in male adults) to induce bone metabolism disturbances in rats (Broulik et al. 2010). In animal models there may be a cause and effect relationship between alcohol consumption and osteonecrosis of the femoral head (Hirota et al. 1993), but for osteonecrosis to develop in humans, the alcohol exposure threshold is approximately 150 l of 100% ethanol, at a consumption rate of 400 ml or more of absolute ethanol weekly (Cruess 1986; Jones 1994). Therefore, we believe that it is difficult to compare rabbit, rat and human studies with regard to the quantity of alcohol used to induce bone disturbances. Bearing this in mind, we suggest that the lack of osteonecrotic lesions in both LDA and HDA groups might be linked to the fact that the doses of alcohol ingested over 4 weeks was too low a dose of alcohol. We believe that the authors need to justify and discuss their choice of dosage. In addition, the short duration of alcohol treatment (4 weeks) is surprising. The authors do not give an explanation for this duration of alcohol treatment. In comparison, alcohol was administered intragastrically to rabbits for one to 6 months, and changes in lipid metabolism were observed only after 2–3 months of treatment (Wang et al. 2003). In another study, mice were treated with alcohol daily for up to 10 months and significant modifications of lipid metabolism appeared at 6 months, with significant increase in largest fat cell size detectable at 10 months (Wang et al. 2008). Finally, animals were treated with alcohol for 4 weeks, and the results were observed at 6 weeks after the beginning of the treatment, therefore after 2 weeks of withdrawal. The authors do not discuss this aspect of the protocol and its relevance in the aetiology of induced osteonecrosis. A possible explanation is to mimic the clinically relevant symptoms of alcohol withdrawal (Schuckit 2009). However, enlightenment about the impact of this withdrawal on osteonecrosis development would be welcome. Thus, without affecting the reported results, we suggest that the findings are difficult to interpret based upon current understanding of alcohol effects on bone metabolism and that there are protocol-related issues, including dose, duration and eventual withdrawal, which might account for the observations reported.

Mesenchymal Cells in the Treatment of Focal Brain Injury Induced by Venous Circulation Disturbances in Rats

We studied the efficiency of bone marrow multipotent mesenchymal stem cells for correction of neurological deficit in rats with experimental sustained focal brain injury caused by venous outflow disturbances. It was found that neurological deficit in animals with transplantation of multipotent mesenchymal stromal cells decreased by 54-75% (vs. 14-17% in the control group) by day 21. The efficiency of mesenchymal stromal cell transplantation (intravenous or local) on day 1 of postoperation period was significantly higher than on day 7. This manifested in more pronounced decrease in the severity of neurological disorders (according to modified neurological severity score) and more rapid recovery of autonomic feeding. Moreover, transplantation of multipotent mesenchymal stromal cells on day 1 abolished the need in antiedematous therapy, while in animals receiving cell therapy on day 7, administration of glucocorticoids was necessary. Comparison of two regimens of cell administration at early terms revealed no advantages of local transplantation of multipotent mesenchymal stromal cells over intravenous injection.

Effect of recombinant interleukin-10 on some haematological and biochemical parameters in a rat endotoxaemic model

Recombinant interleukin-10 (rIL10) has been found to suppress the synthesis of tumour necrosis factor (TNF), interleukin-1 (IL-1), interleukin-6 (IL-6) and tissue factor and to improve survival from experimental sepsis. The aim of this study was to evaluate the protective effect of rIL-10 on lipopolysaccharide-(LPS-) induced haematological and biochemical disturbances in rats. In the present study, 40 rats were used and divided equally into four groups. Group 1 (control group, C) was treated with 0.9% saline. Group 2: LPS was injected intravenously (1.6 mg/100 g), Group 3 received rIL10 treatment (125 μg/kg) 2 min before 0.9% saline injection, Group 4 received rIL10 treatment 2 min before endotoxin treatment. When compared with the controls, platelet count, leukocyte count (with a marked neutrophilia and lymphopenia) and fibrinogen were decreased, while activated partial thromboplastin time (APTT) and prothrombin time (PT) were prolonged in the endotoxaemic rats. In addition, LPS caused statistically significant increases in plasma aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities as well as creatinine, cholesterol and triglyceride concentrations, while it caused a statistically significant decrease in glucose, total protein and albumin levels as compared to the control group. On the other hand, rIL10 significantly suppressed disturbances in the haematological and biochemical parameters associated with endotoxaemia. As a result, rIL10 may be efficacious in preventing haematological disorders, tissue damage and changes in lipid, protein and carbohydrate metabolism in endotoxaemia.